Product Catalog 2013 (PDF) - EUROIMMUN US
Product Catalog 2013 (PDF) - EUROIMMUN US
Product Catalog 2013 (PDF) - EUROIMMUN US
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<strong>EUROIMMUN</strong> Medizinische<br />
Labordiagnostika<br />
AG<br />
<strong>Product</strong> <strong>Catalog</strong>ue<br />
Diagnostics for the Determination of Autoantibodies,<br />
for Infectious Serology and Allergology<br />
Indirect Immunofl uorescence — ELISA — RIA — Westernblot<br />
EUROASSAY — EUROLINE — EUROPL<strong>US</strong> — EUROArray<br />
<strong>2013</strong>
<strong>EUROIMMUN</strong> Medizinische<br />
Labordiagnostika<br />
AG<br />
— 2 —<br />
Table of Contents<br />
<strong>EUROIMMUN</strong> – Company Profile ...............................................................................................................................................3<br />
Techniques for the Serological Investigation of Antibodies .....................................................................................................5<br />
Indirect Immunofluorescence: An Easy and Modern Method .......................................................................................................................... 6<br />
The Indirect Immunofluorescence Test, Performed Using the TITERPLANE Technique ........................................................................... 10<br />
Recommended Serum Dilutions for Indirect Immunofluorescence ............................................................................................................... 12<br />
Diagnostically Relevant Systemic Autoantibodies ........................................................................................................................................... 16<br />
Organ-/Tissue-Specific Autoantibodies ............................................................................................................................................................. 17.<br />
Antibodies for Infectious Serology .................................................................................................................................................................... 18<br />
Antibodies for Allergology ................................................................................................................................................................................. 19<br />
<strong>EUROIMMUN</strong> Microplate ELISA ........................................................................................................................................................................ 20<br />
ELISA Automation using the <strong>EUROIMMUN</strong> Analyzer I ................................................................................................................................... 22<br />
Incubating the Microplate ELISA ....................................................................................................................................................................... 23<br />
EUROASSAY: Line Blots in TITERPLANETechnique Format ....................................................................................................................... 24<br />
Incubating the EUROASSAY (TITERPLANETechnique) ................................................................................................................................ 25<br />
The EUROLINE: A New Technique for Extensive Antibody Profiles .............................................................................................................. 26<br />
Modern Automation of Immunoassays ............................................................................................................................................................ 27.<br />
Westernblots/EUROLINE-WB: Reliable Differentiation of Antibodies Present ............................................................................................... 28<br />
Incubating the EUROLINE/Westernblot/EUROLINE-WB ................................................................................................................................... 29<br />
The EUROArray: DNA Microarray Test Systems for Diagnostics (IVD).......................................................................................................... 30<br />
Performance and Evaluation of the EUROArray Test ...................................................................................................................................... 31<br />
<strong>EUROIMMUN</strong> Radioimmunoassays (RIA/IRMA) .............................................................................................................................................. 32<br />
<strong>EUROIMMUN</strong> <strong>Product</strong>s for the Determination of Autoantibodies ...........................................................................................33<br />
Autoantibodies against Cell Nuclei (ANA) ........................................................................................................................................................ 34<br />
Autoantibodies against Double-Stranded DNA (dsDNA) ................................................................................................................................ 37.<br />
Autoantibodies against CCP and Sa .................................................................................................................................................................. 38<br />
Autoantibodies against Mitochondria (AMA) ................................................................................................................................................... 39<br />
Autoantibodies against Liver Antigens ............................................................................................................................................................. 40<br />
Autoantibodies against Thyroid Gland Antigens / Antigen Detections .......................................................................................................... 42<br />
Autoantikörper against Antigens of the Skin ................................................................................................................................................... 43<br />
Autoantibodies against Neuronal Antigens ...................................................................................................................................................... 44<br />
Autoantibodies against Islet Cell Antigens ....................................................................................................................................................... 46<br />
Autoantibodies against Parietal Cells (PCA) ..................................................................................................................................................... 47.<br />
Autoantibodies against Granulocyte Cytoplasm (cANCA/pANCA) ................................................................................................................. 48<br />
Autoantibodies against CIBD-relevant Antigens .............................................................................................................................................. 49<br />
Autoantibodies against Phospholipase A 2 Receptor (PLA2R) ......................................................................................................................... 51<br />
Antibodies against Endomysium and Gliadin .................................................................................................................................................. 52<br />
<strong>EUROIMMUN</strong> <strong>Product</strong>s for Infectious Serology ......................................................................................................................53<br />
Antibodies against Borrelia ................................................................................................................................................................................ 54<br />
Antibodies against Epstein-Barr Virus (EBV) .................................................................................................................................................... 56<br />
Antibodies against Helicobacter Pylori ............................................................................................................................................................. 58<br />
Antibodies against Herpes Simplex Virus (HSV) ............................................................................................................................................. 59<br />
Antibodies against Chlamydia ........................................................................................................................................................................... 60<br />
Antibodies against Emerging Viruses ............................................................................................................................................................... 61<br />
BIOCHIP Mosaics for Infectious Serology ..................................................................................................................................................... 62<br />
Additional Reagents for the Determination of Acute Infections ..................................................................................................................... 64<br />
<strong>EUROIMMUN</strong> <strong>Product</strong>s for Allergology ...................................................................................................................................65<br />
Order Information and <strong>Product</strong> Data .......................................................................................................................................64<br />
Fluorescence-Labelled Antibodies: Fluorescein (FITC) for <strong>EUROIMMUN</strong> IIFT ............................................................................................... 69<br />
Controls for <strong>EUROIMMUN</strong> IIFT: Organ-Specific Autoantibodies .................................................................................................................... 7.0<br />
Controls for <strong>EUROIMMUN</strong> IIFT: Systemic Autoantibodies ............................................................................................................................. 7.2<br />
Controls for <strong>EUROIMMUN</strong> IIFT: Infectious Serology ....................................................................................................................................... 7.3<br />
Controls for <strong>EUROIMMUN</strong> IIFT: Determination of Further Antibodies .......................................................................................................... 7.9<br />
EUROASSAY for the Determination of Autoantibodies (Test Systems) ........................................................................................................ 80<br />
EUROASSAY for Allergology (Test Systems) ................................................................................................................................................... 81<br />
EUROLINE for the Determination of Autoantibodies (Test Systems) ............................................................................................................. 82<br />
EUROLINE for Infectious Serology (Test Systems) .......................................................................................................................................... 83<br />
EUROLINE for Allergology (Test Systems) ....................................................................................................................................................... 83<br />
Westernblot/EUROLINE-WB for the Determination of Autoantibodies (Test Systems) ................................................................................ 87.<br />
Westernblot/EUROLINE-WB for Infectious Serology (Test Systems) ............................................................................................................. 87.<br />
Microplate ELISA for the Determination of Autoantibodies (Test Systems) ................................................................................................. 89<br />
EUROArray for Molecular Genetic Determinations (Test Systems) ............................................................................................................... 92<br />
Radioimmunoassay (RIA) for the Determination of Autoantibodies / Autoantigens / Hormone Determination (Test Systems) .............. 93<br />
Microplate ELISA for Infectious Serology (Test Systems) .............................................................................................................................. 94<br />
Microplate ELISA for the Determination of Antibodies against Other Antigens (Test Systems) ................................................................100<br />
Microplate ELISA for the Determination of Hormones and Proteins (Test Systems) ..................................................................................100<br />
Allercoat 6 System .........................................................................................................................................................................................101<br />
Diagnostics for Indirect Immunofluorescence: Organ-Specific Autoantibodies ..........................................................................................121<br />
Diagnostics for Indirect Immunofluorescence: Systemic Autoantibodies ...................................................................................................130<br />
Diagnostics for Indirect Immunofluorescence: Infectious Serology .............................................................................................................137.<br />
Diagnostics for Indirect Immunofluorescence: Other Antigens ....................................................................................................................146<br />
<strong>Product</strong>s for EUROPattern ................................................................................................................................................................................147.<br />
<strong>Product</strong>s for EUROLabLiquidHandler ..............................................................................................................................................................148<br />
Reagents and Other Items for <strong>EUROIMMUN</strong> Westernblot and EUROLINE ..................................................................................................149<br />
Further Reagents for <strong>EUROIMMUN</strong> IIFT ..........................................................................................................................................................149<br />
Other Items for <strong>EUROIMMUN</strong> IIFT ...................................................................................................................................................................150<br />
General Delivery Conditions .............................................................................................................................................................................151<br />
Index ......................................................................................................................................................................................152
<strong>EUROIMMUN</strong> Medizinische<br />
Labordiagnostika<br />
AG<br />
Company site groß groenau<br />
Company branch Rennersdorf<br />
<strong>EUROIMMUN</strong> Ag – COMPANY PROFILE<br />
Company headquarters Luebeck Company branch Dassow<br />
Company branch Pegnitz<br />
<strong>EUROIMMUN</strong> worldwide<br />
<strong>EUROIMMUN</strong> was founded in September 1987. and today has its headquarters in Luebeck, Germany. German<br />
branches are situated in gross groenau near Luebeck (Schleswig Holstein), Dassow (Mecklenburg-Western<br />
Pomerania), Rennersdorf (Upper Lusatia, Saxony) and in Pegnitz (Upper Franconia, Bavaria). Further<br />
<strong>EUROIMMUN</strong> AG subsidiaries can be found in Canada (Mississauga), China (Beijing, Hangzhou), great Britain<br />
(Wimbledon), Italy (Padua), Poland (Wroc ław), Switzerland (Lucerne), Singapore, South Africa (Cape town),<br />
Brazil (Porto Alegre), Turkey (Istanbul) and the <strong>US</strong>A (New Jersey). At present <strong>EUROIMMUN</strong> has 944 employees<br />
in Germany, 1311 worldwide. The company is ISO-certified (EN ISO 9001:2008, EN ISO 13485:2003/CMDCAS).<br />
<strong>EUROIMMUN</strong> produces reagents for medical laboratory diagnostics. In the foreground are test systems for<br />
the determination of various antibodies in patient serum in the diagnosis of autoimmune diseases, infectious<br />
diseases and allergies.<br />
The test methods employed are predominantly indirect immunofluorescence, microplate ELISA, various blot<br />
techniques (Westernblot, EUROASSAY, EUROLINE, EUROLINEWB) and all molecular biology techniques. The<br />
company is based on worldwide-patented state-of-the-art production methods and microanalysis techniques<br />
and is one of the world’s leading manu facturers of medical laboratory diagnostics.<br />
The BIOChIPs are one of <strong>EUROIMMUN</strong>’s many inventions: paper-thin sheets of glass are coated with cells<br />
or tissue sections and then cut automatically into millimetre-sized fragments which are subsequently glued<br />
onto slides using a fully automated device. This BIOChIP technology allows extreme miniaturization and<br />
standardization of immun bio chemical analyses. With BIOChIP Mosaics made from 30 or more different<br />
organ sections, cell substrates or defined antigens (EUROPL<strong>US</strong>) only mini mum incubation efforts are<br />
necessary to obtain a detailed antibody profile.<br />
In <strong>EUROIMMUN</strong> enzyme immunoassays (ELISA) defined antigens, purified using state-of-the-art biotechnological<br />
processes, are employed as the antigen substrate. Some of these antigens are synthesized<br />
in the company’s molecular biology laboratories. <strong>EUROIMMUN</strong> ELISA are characterized by their excellent<br />
stability, simple handling and short incubation times, and they are ideal for automated use. All reagents are<br />
delivered ready-to-use and are exchangeable between different lots. <strong>EUROIMMUN</strong> offers the largest and most<br />
differentiated arsenal of enzyme immunoassays worldwide for the diagnosis of autoimmune and infectious<br />
diseases.<br />
— 3 —
<strong>EUROIMMUN</strong> Medizinische<br />
Labordiagnostika<br />
AG<br />
The innovative procedures EUROASSAY and EUROLINE developed by <strong>EUROIMMUN</strong> follow the same test<br />
principle as the ELISA methods, with the use of the BIOCHIP technology. At <strong>EUROIMMUN</strong> purified antigens<br />
are printed in parallel lines at defined positions on membrane strips. Following incubation, users can evaluate<br />
results visually without additional equipment. These reagents allow in particular the differentiation of<br />
antibodies that are not clearly defined microscopically by indirect immunofluorescence. EUROASSAY and<br />
EUROLINE are also em ployed in laboratories where no sophisticated laboratory instruments are available.<br />
<strong>EUROIMMUN</strong> produces an extensive range of Westernblot strip test systems and corres ponding reagents<br />
for confirmation of positive fluorescence and ELISA results as well as clarification of difficult-to-interpret<br />
results in autoimmune diagnostics, infectious serology and allergology. Refined electrophoretical processes<br />
have been developed to allow the precise separation of diagnostically relevant proteins from one another.<br />
Lot-specific evalu ation templates are produced for the evaluation of band patterns. The program ”EURO<br />
LineScan“ enables fully automated evaluation of membrane-based test systems and simplifies the archiving<br />
of results with large sample series.<br />
One of the company’s main strengths is its technical expertise. This encompasses not just the manufacture<br />
and sale of medical laboratory diagnostics, but also the diagnostic application of the products in a reference<br />
laboratory which provides highly differential diagnostics. This reference laboratory has set standards<br />
in Germany, and worldwide is unequalled in the whole field of autoimmune diagnostics. The diagnostic<br />
spectrum of the laboratory also covers the areas of infectious serology and serological allergy diagnostics.<br />
The reference laboratory receives hundreds of serum samples daily from all over Germany as well as from<br />
many other countries. It helps <strong>EUROIMMUN</strong> customers to secure their results: a large proportion of serum<br />
samples sent to <strong>EUROIMMUN</strong> for evaluation are analysed free of charge in order to maintain high standards<br />
in the laboratories of <strong>EUROIMMUN</strong> customers. Customers can obtain further technical information from<br />
experienced scientists in the company, with whom they can also discuss serological problem cases. The<br />
“Institute for Quality Assurance”, an institution newly founded by <strong>EUROIMMUN</strong>, organises unbiased quality<br />
assessments and provides advice in the area of quality management. Moreover <strong>EUROIMMUN</strong> has established<br />
the “Institute for experimental Immunology”, which is engaged in basic research.<br />
In October 2012 the <strong>EUROIMMUN</strong> workforce included 170 university and college graduates, among these<br />
biologists, biochemists, chemists, engineers and medical doctors (48 of them holding a doctor’s degree).<br />
Medical technicians are particularly strongly represented with 111 people, corresponding to <strong>EUROIMMUN</strong>’s<br />
activities, as well as biology/chemistry laboratory technicians (81). At present the company is training 51<br />
young people as biology laboratory assistants, industrial clerks, office communication clerks, IT specialists,<br />
IT clerks, electronic system technicians, electronic technicians for devices and systems, industrial mechanics,<br />
cooks, computer scientists and business economists (dual system). At <strong>EUROIMMUN</strong> great value is placed on<br />
advising customers and prospective customers in a factual, technical and commercially restrained manner<br />
and fully supporting them in the use of our diagnostically demanding products. <strong>EUROIMMUN</strong> products are<br />
backed by an energetic and competent sales force, qualified information material, didactic test instructions<br />
and scientifically based, but nevertheless understandable advertisements in technical journals. Advertising<br />
material is produced in-house using the latest desktop publishing methods, right up until the fully digitalised<br />
ready-for-exposure documents. The most important publications and posters are translated into many<br />
languages. <strong>EUROIMMUN</strong> has set up an informative homepage on the internet (www.euroimmun.com) which<br />
is visited extensively internationally.<br />
Over 3,000 laboratories worldwide use <strong>EUROIMMUN</strong> diagnostics. 400 of these are in Ger many. The company’s<br />
development is shaped by continuous growth. Although the diagnostic market in Germany stagnated and<br />
has become particularly strongly competitive, EURO IMMUN has been able to continue its strong expansion.<br />
The company is achieving an ever increasing independence from the German market, since more and more<br />
products are sold abroad. With their quality and standardization, <strong>EUROIMMUN</strong> products are capturing the<br />
leading position in the world.<br />
— 4 —
<strong>EUROIMMUN</strong> Medizinische<br />
Labordiagnostika<br />
AG<br />
INvESTIgATION TEChNIqUES<br />
— 5 —<br />
Investigation<br />
Techniques
<strong>EUROIMMUN</strong> Medizinische<br />
Labordiagnostika<br />
AG<br />
Investigation<br />
Techniques<br />
— 6 —<br />
cell with<br />
antigen<br />
specific human<br />
antibody<br />
FITC<br />
Indirect Immunofluorescence: An Easy and Modern Method<br />
FITC<br />
Pattern homog. AntidsDNA?<br />
Anti-Histones?<br />
Pattern nucleolar. Anti-<br />
PM-Scl?<br />
FITC-labelled antihuman<br />
antibody<br />
Pattern fine-granular.<br />
Anti-SS-A? Anti-SS-B?<br />
Pattern cytoplasmic.<br />
AMA M2?<br />
Differentiation of antibodies using HEp-2 cells.<br />
tissue sections<br />
antigen dots<br />
BIOCHIP Technology and Mosaics.<br />
culture cells<br />
transf.<br />
cells<br />
Principle of the Test<br />
• For the determination of autoantibodies or antibodies against infectious agents,<br />
cells, tissue sections or purified, biochemically characterized substances are<br />
used as antigen substrates.<br />
• If the sample is positive, specific antibodies in the diluted serum sample attach<br />
to the antigens coupled to a solid phase.<br />
• In a second step, the attached antibodies are stained with fluorescein-labelled<br />
anti-human antibodies and visualized with the fluorescence microscope.<br />
• Positive samples can be titrated in steps. The most suitable titration interval<br />
is provided by the dilution factor 3.162 (square root of 10). In this way, every<br />
second step represents in its denominator an integral power of 10 (1 : 10, 1 : 32,<br />
1 : 100, 1 : 320, 1 : 1000, 1 : 3200, 1 : 10000 etc.).<br />
Indirect Immunofluorescence: A Standardized Technique for the<br />
Determination of Autoantibodies and Antibodies against Infectious<br />
Agents<br />
• High specificity: positive and negative samples produce a large difference in<br />
signal strength. Each bound antibody shows a typical fluorescence pattern<br />
depending on the location of the individual antigens.<br />
• The entire antigen spectrum of the original subtrate is available, thus allowing<br />
the detection of a large number of antibodies and achieving a higher detection<br />
rate.<br />
• Immunofluorescence enables simultaneous detection of antibodies against<br />
several biochemically different antigens on one single biological substrate.<br />
• The indirect immunofluorescence test is the analytical method of choice when<br />
it would be too difficult or too complicated to prepare the test antigens individually<br />
for enzyme immunoassays.<br />
<strong>EUROIMMUN</strong>’s Innovations for the Standardization and Modernization<br />
of Indirect Immunofluorescence<br />
• Activation technique: physically or chemically activated cover glasses are coated<br />
with cultured cells or tissue sections. Frozen tissue sections are fixed to the<br />
glass surface by covalent bonding, increasing adhesion more than 100 times<br />
and thus preventing the substrates from being detached.<br />
• BIOCHIP Technology: cover glasses coated with biological substrates are cut<br />
into millimetre-sized fragments (BIOCHIPs) on a machine. This makes it possible<br />
to obtain ten or more first-class preparations of homogeneous quality per tissue<br />
section, in the case of cultured cell substrates even several thousands.<br />
• BIOCHIP Mosaics: using several BIOCHIPs coated with different substrates<br />
side by side on one and the same reaction field, antibodies against various<br />
organs or infectious agents can be investigated simultaneously. Detailed antibody<br />
profiles can thus be established with comparatively little effort, allowing<br />
the reciprocal determination of the results on different substrates.<br />
• TITERPLANE Technique: samples or reagents are applied to the reaction fields<br />
of a reagent tray. The BIOCHIP Slides are then placed into the recesses of the<br />
reagent tray, where all BIOCHIPs come into contact with the fluids, and the<br />
individual reactions commence simultaneously. As the fluids are confined in a<br />
closed space, there is no need for the use of a conventional „humidity chamber“.
<strong>EUROIMMUN</strong> Medizinische<br />
Labordiagnostika<br />
AG<br />
Indirect Immunofluorescence: An Easy and Modern Method<br />
Chemically Activated Cover Glasses for Histochemistry<br />
• For diagnostics of organ-specific or tissue-specific autoantibodies frozen tissue<br />
sections of various organs are used. However, formerly, the morphology of<br />
tissues suffered during incubation in aqueous medium, tissue parts occasionally<br />
became detached from slides, and the interpretation of results was difficult.<br />
• Using the activation technique for the first time in histology, we have applied<br />
solid phase techniques. Firstly, the surface of cover glasses is coated with<br />
spontaneously reactive aldehyde groups. In a second step, the tissue sections<br />
are applied to the chemically activated cover glasses (Stöcker, W: European<br />
Patent No. 0 117 262; U.S. Patent No. 4,647,543). Free amino groups of the tissue<br />
sections, especially of the hydroxy lysine contained in the collagen, bind to the<br />
carrier material by covalent bonding.<br />
• This results in an increased adhesion of frozen tissue sections more than a<br />
hundredfold and prevents them from being detached during incubation.<br />
Furthermore, in some cases the activation technique results in a significantly<br />
better conservation of tissue structures, especially in organs which previously<br />
exhibited a generally low level of adhesion. Therefore, the tests can be evaluated<br />
with considerably greater confidence.<br />
Determination of Low-Avidity Antibodies<br />
• An alternative principle for the serological diagnosis of fresh infections has been<br />
established by investigating the antibody avidity.<br />
• The first reaction of the immune system following an infection is the formation<br />
of low-avidity antibodies. As the infection proceeds, increasingly antigen-adapted<br />
IgG is formed, and avidity grows. As long as high-avidity IgG is not yet detected<br />
in the serum, it can be assumed that the infection is still in an early stage.<br />
• To identify low-avidity antibodies in a patient’s serum, two immunofluorescence<br />
tests are performed in parallel: one test is carried out in the conventional way,<br />
the other one includes urea treatment between incubations with patient’s serum<br />
and per oxidase-labelled anti-human IgG, resulting in the detachment of lowavidity<br />
antibodies from the antigens.<br />
• Low-avidity antibodies are present if the fluorescence intensity is significantly<br />
reduced (two intensity levels ore more) by urea treatment.<br />
• The following test kits for avidity determination are available: Toxoplasma<br />
gondii, Rubella virus, West-Nile virus, EBV-CA, EBV-EA, VZV, CMV.<br />
EUROPL<strong>US</strong> System: Combination of conventional immunofluorescence<br />
substrates and monospecific tests<br />
• In EUROPL<strong>US</strong> immunofluorescence tests antibody detection is performed using<br />
both tissue sections/cell substrates and monospecifically reacting antigens.<br />
• Antibodies detected in IFT screening tests can therefore be differentiated or<br />
confirmed with one and the same reaction field. In some cases, the antigens<br />
help to extend the antigen spectrum, offering a wider range for screening.<br />
• BIOCHIPs coated with purified or recombinant antigens are used as monospecific<br />
substrates.<br />
• In case of a positive result the antigens fluoresce green in defined areas under<br />
the microscope.<br />
• In some EUROPL<strong>US</strong> test systems several different antigens are coated on<br />
one BIOCHIP in separate antigen rows. In this manner, several monospecific<br />
analyses can be performed using a single BIOCHIP.<br />
• Available EUROPL<strong>US</strong> substrates: MPO, PR3, gliadin GAF-3X, OspC, VlsE,<br />
Plasmodium falciparum und vivax, gp125, p19.<br />
Aminoethylaminoproyltrimethoxysilane<br />
NH 2<br />
(CH 2)2<br />
NH<br />
(CH 2)3<br />
H3CO Si OCH3 OCH 3<br />
OH<br />
glass<br />
glutardialdehyde<br />
HC O<br />
(CH 2)3<br />
HC O<br />
frozen tissue section<br />
HC O<br />
- 3 CH3OH<br />
NH2 - H2O N<br />
- H2O<br />
(CH2)2 (CH2)2 NH<br />
(CH 2)3<br />
Si O<br />
O<br />
Si O<br />
Fixation of frozen tissue sections to glass surfaces<br />
by covalent bonding.<br />
(CH 2)3<br />
(CH 2)3<br />
low-avide Ab against EBV-CA<br />
high-avide Ab against EBV-CA<br />
without urea with urea<br />
NH 2<br />
HC<br />
NH<br />
O<br />
frozen tissue section<br />
N<br />
HC<br />
(CH 2)3<br />
HC<br />
N<br />
(CH 2)2<br />
NH<br />
(CH 2)3<br />
Si O<br />
EUROPL<strong>US</strong>: BIOCHIP combination of tissue<br />
sections / cell substrates (left) and purified<br />
antigens (right).<br />
O<br />
— 7. —<br />
Investigation<br />
Techniques
<strong>EUROIMMUN</strong> Medizinische<br />
Labordiagnostika<br />
AG<br />
Investigation<br />
Techniques<br />
— 8 —<br />
Indirect Immunofluorescence: An Easy and Modern Method<br />
IF Sprinter: Reliable results for small and medium throughput<br />
• Safe and convenient: Fully automated processing of immunofluorescence tests,<br />
from the dilution and dispensing of samples to the incubation and washing of<br />
microscope slides.<br />
• User-friendly: Sample identification by automatic scanning of barcodes when<br />
racks are inserted into the system.<br />
• Quick and reliable results: The washing of slides by flooding ensures short<br />
processing times and clear immunofluorescence signals.<br />
• Smooth laboratory routine: The connection to EUROLabOffice (optional) offers<br />
unique ways to optimise processes in serology, e. g. automatic generation of<br />
worklists.<br />
• 96 primary tubes (10 - 13 mm tube diameter).<br />
• 12 controls and 8 reagents (customised <strong>EUROIMMUN</strong> racks).<br />
• Up to 20 slides (depending on the system configuration).<br />
• 192 dilution positions.<br />
Sprinter XL: Fast IFT automation for medium and high throughput<br />
- also available for ELISA<br />
• Consolidation: fully automated processing of immunofluorescence tests and<br />
ELISA on one system / in one run.<br />
• Quick results: 4 washable needles and 2 arms allow efficient and optimised time<br />
management of worklists.<br />
• Flexibility: various system configurations available to suit different laboratory<br />
needs.<br />
• Quality and security: modern washing technology for brilliant fluorescence, clot<br />
detection, liquid level detection and precise pipetting.<br />
• User-friendly software and system operation: only four steps to start a worklist.<br />
• 160 (or 240) primary tubes (10 - 16 mm tube diameter).<br />
• 64 (49) controls / calibrators, 20 (12) secondary reagents, 6 samples buffers and<br />
4 wash buffers.<br />
• Up to 30 slides / 6 microplates.<br />
• 162 screening dilutions / 192 titer dilutions.<br />
AP16 IF Plus by DAS: Automation Solution for all <strong>EUROIMMUN</strong><br />
Immunofluorescence Tests<br />
• Various validated parameters. Slide definitions and test files available.<br />
• CE conformity for device/test system combination.<br />
• Capacity: 16 slides, 80 samples, 200 dilutions.<br />
• Programmable for 8 methods per run.<br />
• 12 dilution series freely programmable.<br />
• Automated sample dilution, sample and reagent dispension, incubation and<br />
washing of slides.<br />
• Laboratory software interface.<br />
• The incubation protocol for result documentation is automatically created from<br />
the worklist.<br />
• Barcode reader available on request.<br />
• Very simple operation.
<strong>EUROIMMUN</strong> Medizinische<br />
Labordiagnostika<br />
AG<br />
Indirect Immunofluorescence: An Easy and Modern Method<br />
EUROPattern: Computer-Aided Immunofluorescence Microscopy<br />
(CAIFM)<br />
• EUROPattern is a high-performance automation solution designed by EURO-<br />
IMMUN for the evaluation of immunofluorescence slides in autoimmune<br />
diagnostics.<br />
• IIFT patterns and corresponding titers are automatically identified and given for<br />
each patient individually (including mixed patterns).<br />
• Validation and export to LIS are carried out at a mouse click using EUROLabOffice<br />
(ELO).<br />
• Automated identification of slides via matrix codes.<br />
• Automated processing of up to 500 analysis positions in succession.<br />
• 3D hand control for manual operation.<br />
• Controlled LED (> 50,000 hrs of constant light flux).<br />
• No dark room required.<br />
EUROPattern Software<br />
• Automated image acquisition of tissue sections.<br />
• Evaluation of results at the screen: manual / visual or automated.<br />
• Optionally with automated pattern recognition (HEp-2 / HEp-20-10), including<br />
titer (also for mixed patterns).<br />
• Result report per patient (consolidation of single results).<br />
• Archiving of images and results.<br />
• Access to patient history.<br />
• Selection of follow-up analyses.<br />
• Lists of positives / negatives.<br />
Fluorescence Microscope EUROStar III Plus and <strong>EUROIMMUN</strong> cLED<br />
• EUROStar III Plus is specifically tailored to the requirements of indirect immunofluorescence.<br />
The conventional complex illumination fittings have been replaced<br />
by the stunningly simple EUROStar Bluelight system.<br />
• The LED has a life expectancy of 50,000 hours – which is 500 times longer than<br />
a mercury vapour lamp. Its light intensity is maintained at a constant level by<br />
electronic regulation. Thus, the microscope requires almost no maintenance.<br />
• Our technicians regularly check the light intensity of your EUROStar III Plus and<br />
issue a certificate to support your quality management system.<br />
• LEDs require only a tenth of the electrical power of a 50-watt HBO lamp, at a<br />
comparable brightness. EUROStar Bluelight provides instant full light output<br />
every time after being switched on. It does not emit any ultraviolet radiation and<br />
is explosion-proof.<br />
• Switching between the camera and the eyepieces is unnecessary due to the<br />
convenient 50/50 beam splitter.<br />
• With its halogen transmitted-light source, EUROStar III Plus is suited for brightfield,<br />
darkfield and, optionally, for phase contrast microscopy.<br />
• With the <strong>EUROIMMUN</strong> cLED, the EUROStar Bluelight technology is also available<br />
as a separate component to upgrade other microscope types.<br />
Pos.<br />
Neg.<br />
Borderline<br />
?<br />
Pos.<br />
Neg.<br />
Borderline<br />
?<br />
Repeat<br />
Pos.<br />
Neg.<br />
Borderline<br />
?<br />
Computer result<br />
+++ Centromeres 1:3,200<br />
Visual microscopic result<br />
++++ Centromeres 1:10,000<br />
Addition:<br />
ENA EUROPL<strong>US</strong> neg.<br />
Final result<br />
+++ Centromeres 1:3,200<br />
Unofficial remark<br />
Further<br />
diagnostics<br />
Macros<br />
98%<br />
Delete all X Reset<br />
Homogeneous pattern<br />
Granular pattern<br />
Nucleolar pattern<br />
Centromeres<br />
Nuclear dots<br />
Mitosis pattern<br />
Cytoplasmic pattern<br />
Negative<br />
Nuclear membrane<br />
Ku<br />
PCNA<br />
Mitosin (CENP F)<br />
Coilin (Few nuclear dots)<br />
Spindle apparatus<br />
Centrioles<br />
Jo-1<br />
Other<br />
Verify synthesis<br />
15/28<br />
Serum No.: 127<br />
1:3,200 94%<br />
?<br />
— 9 —<br />
Investigation<br />
Techniques
<strong>EUROIMMUN</strong> Medizinische<br />
Labordiagnostika<br />
AG<br />
Investigation<br />
Techniques<br />
— 10 —<br />
The Indirect Immunofluorescence Test, Performed Using the<br />
TITERPLANE Technique<br />
(Reaction fields 5 x 5 mm)<br />
The TITERPLANE Technique was developed by <strong>EUROIMMUN</strong> in order to standardize immunological analyses:<br />
Samples or labelled antibodies are applied to the reaction fields of a reagent tray. The BIOChIP Slides are then<br />
placed into the recesses of the reagent tray, where all BIOCHIPs of the slide come into contact with the fluids,<br />
and the individual reactions commence simultaneously. Position and height of the droplets are exactly defined<br />
by the geometry of the system. As the fluids are confined in a closed space, there is no need for the use of a<br />
conventional ”humidity chamber”. It is possible to incubate any number of samples next to each other and<br />
simultaneously under identical conditions.<br />
Prepare: Check the reagent tray: Are the reaction fields hydrophiIic and the surrounding coating hydro phobic?<br />
If not, rub with a wet paper towel, using normal household detergent or Extran MA 01 (Merck) if necessary, and<br />
rinse thoroughly with water. For occasional disinfection, immerse for 1 h in 3% Sekusept Extra (Henkel) in water.<br />
Open the individual packets containing the BIOCHIP Slides only after they have reached room temperature. Do<br />
not touch the BIOCHIPs. Mark BIOCHIP Slides as required with a felt pen.<br />
Dilute: Dilute serum samples according to the user’s test protocol. Include positive and negative controls with<br />
every test procedure. Mix control sera before use.<br />
Pipette: Apply 30 µl of diluted serum to each reaction field of the reagent tray, avoiding air bubbles. Transfer all<br />
samples to be tested before starting the incubation (up to 200 droplets). Use a polystyrene pipetting template.<br />
Incubate: Start reactions by fitting the BIOCHIP Slides into the corresponding recesses of the reagent tray.<br />
Ensure that each sample makes contact with its BIOCHIP and that the individual samples do not come into<br />
contact with each other. Incubate for 30 min at room temperature.<br />
Wash: Rinse the BIOCHIP Slides with a flush of PBS-Tween using a beaker, and immerse them immediately<br />
afterwards in a cuvette containing PBS-Tween for at least 5 min.<br />
Pipette: Apply 25 µl of fluorescein-labelled anti–human immunoglobulin (conjugate) onto each reaction field<br />
of a clean reagent tray. Add all drops (reagent for a maximum of 50 slides) before continuing incubation. Use<br />
a stepper pipette. The labelled anti-human serum should be mixed with a pipette before use. To save time,<br />
conjugate can be pipetted onto separate reagent trays during incubation with the diluted serum.<br />
Incubate: Remove one BIOCHIP Slide from the PBS-Tween and within five seconds blot only the back and the<br />
long edges with a paper towel and immediately put the BIOCHIP Slide into the recesses of the reagent tray. Do<br />
not dry the areas between the reaction fields. Check for correct contact between the BIOCHIPs and liquids. Then<br />
continue with the next BIOCHIP Slide. From now on, protect the slides from direct sunlight. Incubate for 30 min<br />
at room temperature.<br />
Wash: Rinse the BIOCHIP Slides with a flush of PBS-Tween using a beaker and put them in a cuvette containing<br />
PBS-Tween for at least 5 min. 10 drops of Evans Blue (150 µl) for each 150 ml phosphate buffer can be added for<br />
counterstaining.<br />
Embed: Place glycerol/PBS onto a cover glass – drops of 10 µl per reaction field. Use a polystyrene embedding<br />
template. Remove one BIOCHIP Slide from the PBS-Tween and dry the back and all four edges as well as the surface<br />
around, but not between, the reaction fields with a paper towel. Put the BIOCHIP Slide, with the BIOCHIPs facing<br />
downwards, onto the prepared cover glass. Check immediately that the cover glass is properly fitted into the<br />
recesses of the slide. Correct the position if necessary. Now proceed in the same way with the next BIOCHIP<br />
Slide.<br />
Evaluate: Read the fluorescence under the microscope.
<strong>EUROIMMUN</strong> Medizinische<br />
Labordiagnostika<br />
AG<br />
The Indirect Immunofluorescence Test, Performed Using the<br />
TITERPLANE Technique<br />
Pipette:<br />
10 µl per field (3 x 3 mm)<br />
30 µl per field (5 x 5 mm)<br />
7.0 µl per field (7. x 9 mm)<br />
Incubate: 30 min<br />
Wash: 1 s flush<br />
5 min cuvette<br />
Pipette:<br />
10 µl per field (3 x 3 mm)<br />
25 µl per field (5 x 5 mm)<br />
65 µl per field (7. x 9 mm)<br />
Incubate: 30 min<br />
Wash: 1 s flush<br />
5 min cuvette<br />
Embed:<br />
10 µl per field (3 x 3 mm)<br />
10 µl per field (5 x 5 mm)<br />
20 µl per field (7. x 9 mm)<br />
Evaluate: fluorescence microscopy<br />
slide<br />
BIOCHIPs<br />
PBS-<br />
Tween<br />
PBS-<br />
Tween<br />
20x<br />
NEOFLUAR<br />
reagent tray<br />
diluted samples<br />
labelled antibody<br />
glycerol/PBS<br />
cover glass<br />
— 11 —<br />
Investigation<br />
Techniques
<strong>EUROIMMUN</strong> Medizinische<br />
Labordiagnostika<br />
AG<br />
Investigation<br />
Techniques<br />
— 12 —<br />
Recommended Serum Dilutions for Indirect Immunofluorescence<br />
– Autoimmunity –<br />
Antibodies against Substrate IgA Igg IgM IgAgM<br />
acetylcholine receptor *skeletal muscle, monkey / heart, monkey 100<br />
actin Hepatitis Mosaic**, VSM47. 10 100 + 1000 100 + 1000<br />
ADH-producing cells nucl. supraopticus & paraventricularis, monkey 10 10<br />
adrenal cortex adrenal gland, monkey 10<br />
alveolar basement membrane lung, monkey / kidney, monkey 10<br />
aquaporin-4 Neurology Mosaic 10 + 100 10 + 100<br />
asialoglycoprotein receptors *Hepatitis Mosaic** 100<br />
basic myelin protein (BMP) cerebellum, monkey / nerves, monkey / intestinal tissue, fetal monkey 10 + 100<br />
bile canaliculi Hepatitis Mosaic** 100<br />
bile duct epithelium Hepatitis Mosaic** 100<br />
BP180 Dermatology Mosaic (EUROPL<strong>US</strong>) 10 10 + 100 10 + 100<br />
BP230 Dermatology Mosaic (transfected cells) 10 + 100 10 + 100 10 + 100<br />
brain: grey matter cerebellum, monkey / intestinal tissue, monkey 10 + 100 10<br />
brain: white matter cerebellum, monkey / intestinal tissue, monkey 10 + 100 10<br />
cANCA granulocytes (EOH), human / liver, monkey 10 10 + 100 + 1000<br />
cartilage trachea, fetal monkey 10 10<br />
cell nuclei (ANA) HEp-2 cells / liver, monkey 100 + 1000 + 10000 100<br />
CENP-F *HEp-2 cells / liver, monkey 10 + 100 + 1000 100<br />
centromere HEp-2 cells / liver, monkey 100 + 1000 + 10000 100<br />
cerebrum gyrus precentralis, monkey 10 + 100 10<br />
chondroitin sulfate trachea / cartilage, monkey 10 10<br />
collagen type VII oesophagus, monkey / transfected cells 10 + 100 10 + 100<br />
collagenous connective tissue pancreas, monkey 10 10<br />
colon (epithelial cells) intestinal tissue, fetal monkey 10 10<br />
cornea eye, monkey 10<br />
CUZD1 (rPAg1) CIBD Mosaic 10 + 100 10 + 100<br />
cyclin I (PCNA) HEp-2 cells / liver, monkey 10 + 100 + 1000 100<br />
cyclin II (mitosin) HEp-2 cells / liver, monkey 10 + 100 + 1000 100<br />
cytoskeleton HEp-2 cells / liver, monkey 100 10 + 100 + 1000 100<br />
desmoglein 1+3 Dermatology Mosaic (transfected cells) 10 + 100 10 + 100 10 + 100<br />
desmosomes oesophagus, monkey or tongue, monkey 10 + 100 10 + 100 10 + 100<br />
dsDNA Crithidia luciliae 10 10 10 10<br />
dsDNS-bound laktoferrin CIBD Mosaic 10 10<br />
elastin stomach, rat / kidney, rat 100 + 1000<br />
endocardium endocardium, monkey / intestinal tissue, monkey 10<br />
endomysium liver, monkey 10 10<br />
endothelial cells skeletal muscle, monkey / HUVEC 100 100<br />
endplates *skeletal muscle, monkey / heart, monkey 10 + 100<br />
enterocytes intestinal tissue, fetal monkey 10 10<br />
eosinophilic granulocytes granulocytes (EOH) / liver, monkey 10 10 + 100 1<br />
epidermal basement membrane oesophagus, monkey or tongue, monkey 10 + 100 10 + 100<br />
eye muscle eye, monkey 100<br />
filaggrin oesophagus, rat 10 10<br />
GABA-receptor B1 transfected cells 10 + 100<br />
ganglion cells ganglion stellatum, monkey / intestinal tissue, monkey 10<br />
gastric mucosa stomach, monkey 10<br />
gastrin-producing cells stomach (antrum), monkey / stomach (corpus), monkey 10<br />
glandula suprarenalis adrenal gland, monkey 10<br />
gliadin *intestinal tissue, fetal monkey / gliadin dots 10 10<br />
glomerular basement membrane (GBM) kidney, monkey 10 10<br />
glutamate receptor type AMPA transfected cells 10 + 100 10 + 100<br />
glutamate receptor type NMDA transfected cells 10 + 100 10 + 100<br />
glutamic acid decarboxylase (GAD) cerebellum, monkey / pancreas, monkey 10 + 100 10<br />
goblet cells goblet cells (culture) 10 10<br />
Golgi apparatus HEp-2 cells / liver, monkey 100 + 1000 + 10000 100<br />
*) In addition to the preferential analysis or as a plausibility check.<br />
**) Hepatitis Mosaic: liver, monkey / heart, monkey / HEp-2 cells / liver, rat / kidney, rat / stomach, rat.
<strong>EUROIMMUN</strong> Medizinische<br />
Labordiagnostika<br />
AG<br />
Recommended Serum Dilutions for Indirect Immunofluorescence<br />
– Autoimmunity –<br />
Antibodies against Substrate IgA Igg IgM IgAgM<br />
GP2 (rPAg2) CIBD Mosaic 10 10 + 100 + 1000<br />
hair follicle epidermis, monkey 10 10<br />
heart muscle skeletal muscle, monkey / heart, monkey 100 100<br />
heart valves mitral valve, monkey 10 10<br />
histones *HEp-2 cells / liver, monkey 100 + 1000 + 10000 100<br />
Hu cerebellum, monkey / nerves, monkey / intestinal tissue, fetal monkey 10 + 100 10 + 100<br />
hypothalamus nucl. supraopticus & paraventricularis, monkey 10 10<br />
intercalated discs heart, monkey 100 100<br />
intestinal epithelial tissue intestinal tissue, monkey 10 10<br />
intrinsic factor *stomach, monkey / intrinsic factor 10 10<br />
Jo-1 *HEp-2 cells / liver, monkey 10 + 100 + 1000 100<br />
keratin oesophagus, monkey or tongue, monkey 10 10<br />
keratin, RA-associated (filaggrin) oesophagus, rat 10 10 10 10<br />
Ku *HEp-2 cells / liver, monkey 100 + 1000 + 10000 100<br />
labyrinth inner ear, rat or guinea pig 10 10<br />
lacrimal gland (excretory ducts and acini) lacrimal gland, monkey 10 10<br />
lamins HEp-2 cells / liver, monkey 100 + 1000 + 10000 100<br />
lipocytes fat tissue, monkey 10 10<br />
liver membrane (LMA) Hepatitis Mosaic** 100<br />
liver-kidney microsomes (LKM) Hepatitis Mosaic** 100<br />
liver-pancreas antigen (LP) Hepatitis Mosaic** / pancreas, monkey 100<br />
liver-specific protein (LSP) Hepatitis Mosaic** 100<br />
lymphocytes Lymphocytes 10 + 100 10 + 100<br />
lysosomes HEp-2 cells / liver, monkey 100 + 1000 + 10000 100<br />
M2 *Hepatitis Mosaic** 100 + 1000 + 10000 100<br />
M3 *Hepatitis Mosaic** 100 + 1000 + 10000 100<br />
M4 *Hepatitis Mosaic** 100 + 1000 + 10000 100<br />
M5 *Hepatitis Mosaic** 100 + 1000 + 10000 100<br />
M6 *Hepatitis Mosaic** 100 + 1000 + 10000 100<br />
M7. *Hepatitis Mosaic** 100 + 1000 + 10000 100<br />
M8 *Hepatitis Mosaic** 100 + 1000 + 10000 100<br />
M9 *Hepatitis Mosaic** 100 + 1000 + 10000 100<br />
medullated nerves cerebellum, monkey / nerves, monkey 10 + 100<br />
melanocytes retina, monkey 10 10<br />
Mi-2 HEp-2 cells / liver, monkey 100 + 1000 + 10000 100<br />
mitochondria (AMA) kidney, rat / stomach, rat / M2 dots / HEp-2 cells 100 + 1000 + 10000 100<br />
mouth mucosa mouth mucosa 10<br />
myelin cerebellum, monkey / nerves, monkey 100 100<br />
myelin-associated glycoprotein (MAG) cerebellum, monkey / nerves, monkey 10 10 10 + 100 10 + 100<br />
myeloperoxidase (MPO) granulocytes (EOH), human / liver, monkey 10<br />
myocardium skeletal muscle, monkey / heart, monkey 100 100<br />
myolemma skeletal muscle, monkey / heart, monkey 10 10<br />
myosin skeletal muscle, monkey / heart, monkey 100<br />
native collagen pancreas, monkey 10<br />
nerves cerebellum, monkey / nerves, monkey / intestinal tissue, fetal monkey 10 + 100 10 + 100<br />
neuroendothelium cerebellum, monkey / nerves, monkey / intestinal tissue, fetal monkey 10 + 100 10 + 100<br />
neurofilaments cerebellum, monkey / nerves, monkey / intestinal tissue, fetal monkey 10 + 100 10 + 100<br />
NOR HEp-2 cells / liver, monkey 100 + 1000 + 10000 100<br />
nRNP HEp-2 cells / liver, monkey 100 + 1000 + 10000 100<br />
ovary ovary, monkey 10 10<br />
pANCA granulocytes (EOH), human / liver, monkey 10 10 + 100 + 1000<br />
pancreas acini (Crohn’s disease autoantigen) pancreas, monkey 10 + 100 10 + 100<br />
pancreas islets pancreas, monkey (1st step: 18 hours) 10 + 100<br />
pancreas, excretory duct epithelium pancreas, monkey 10 + 100 10 + 100<br />
parathyroid gland parathyroid gland, monkey 10<br />
*) In addition to the preferential analysis or as a plausibility check.<br />
**) Hepatitis Mosaic: liver, monkey / heart, monkey / HEp-2 cells / liver, rat / kidney, rat / stomach, rat.<br />
— 13 —<br />
Investigation<br />
Techniques
<strong>EUROIMMUN</strong> Medizinische<br />
Labordiagnostika<br />
AG<br />
Investigation<br />
Techniques<br />
parietal cells stomach (corpus), monkey 10 10 10 + 100<br />
parotid gland parotid gland, monkey 10 10<br />
peripheral nerves cerebellum, monkey / nerves, monkey / intestinal tissue, fetal monkey 10 + 100 10 + 100<br />
pituitary gland, anterior lobe pituitary gland, monkey 10 10<br />
placenta placenta, human 10<br />
PM-1 HEp-2 cells / liver, monkey 100 + 1000 + 10000 100<br />
prostate prostate, monkey 10<br />
proteinase 3 (PR3) granulocytes (EOH), human / liver, monkey 10 10 + 100 + 1000<br />
Purkinje cell cytoplasm (Yo) cerebellum, monkey 10 + 100 10 + 100<br />
RANA Raji cells / HEp-2 cells 10 10<br />
reticulin intestinal tissue, fetal monkey 10 10 10<br />
retina retina, monkey 10 10<br />
Ri cerebellum, monkey / nerves, monkey / intestinal tissue, fetal monkey 10 + 100 10 + 100<br />
ribosomes HEp-2 cells / liver, monkey 10 + 100 + 1000 100<br />
RNA polymerase HEp-2 cells / liver, monkey 100 + 1000 + 10000 100<br />
RNA HEp-2 cells / liver, monkey 100 + 1000 + 10000 100<br />
salivary glands (acini and excretory ducts) parotid gland, monkey 10 10<br />
sarcolemma skeletal muscle, monkey / heart, monkey 100 10<br />
Scl-7.0 HEp-2 cells / liver, monkey 100 + 1000 + 10000 100<br />
signal recognition particle (SRP) HEp-2 cells / liver, monkey 100 + 1000 + 10000 100<br />
skeletal muscle skeletal muscle, monkey / heart, monkey 100 100<br />
Sm HEp-2 cells / liver, monkey 100 + 1000 + 10000 100<br />
smooth muscles (ASMA) stomach, rat / kidney, rat 100 + 1000 100 + 1000<br />
spermatozoa spermatozoa smear, human 10 10 10<br />
spindle fibers HEp-2 cells / liver, monkey 100 + 1000 + 10000 100<br />
spleen spleen, monkey 10 + 100<br />
SS-A (Ro) *HEp-2 cells / liver, monkey 100 + 1000 + 10000 100<br />
SS-B (La) *HEp-2 cells / liver, monkey 100 + 1000 + 10000 100<br />
ssDNA *HEp-2 cells / liver, monkey 100 + 1000 + 10000 100<br />
striated muscles skeletal muscle, monkey / heart, monkey 100 + 1000 100 + 1000<br />
substantia nigra substantia nigra, monkey 10 + 100<br />
testis testis, monkey 10<br />
thrombocytes (bound antibodies) thrombocyte smear – – –<br />
thrombocytes (free antibodies) thrombocytes, human 10 10 10<br />
thymus thymus, monkey 10 10 10<br />
thyroglobulin thyroid gland, monkey or struma, human/TG 10 + 100 10<br />
thyroid colloid type II thyroid gland, monkey or struma, human 10 10<br />
thyroid microsomes thyroid gland, monkey or struma, human 10 10<br />
trachea trachea, monkey 10 10<br />
tubular basement membrane kidney, monkey 10 10<br />
VGKC (CASPR2 + LGI1) transfected cells 10 + 100 10 + 100<br />
U1-nRNP *HEp-2 cells / liver, monkey 100 + 1000 + 10000 100<br />
vasopressin-producing cells nucl. supraopticus & paraventricularis, monkey 10 10<br />
vestibular organ inner ear, rat or guinea pig 10 10<br />
vimentin *HEp-2 cells / liver, monkey 100 + 1000 + 10000 100<br />
”xANCA” granulocytes (EOH, HCHO) / liver, monkey 10 10 + 100 1<br />
— 14 —<br />
Recommended Serum Dilutions for Indirect Immunofluorescence<br />
– Autoimmunity –<br />
Antibodies against Substrate IgA Igg IgM IgAgM<br />
*) In addition to the preferential analysis or as a plausibility check.
<strong>EUROIMMUN</strong> Medizinische<br />
Labordiagnostika<br />
AG<br />
Recommended Serum Dilutions for Indirect Immunofluorescence<br />
– Infectious Serology –<br />
Antibodies against IgA Igg IgM<br />
Adenovirus (type 3) 10 + 100 10 + 100 10<br />
Bartonella henselae 320 + 1000 100<br />
Bartonella quintana 320 + 1000 100<br />
Bordetella parapertussis 10 + 100 100 + 1000 100 + 1000<br />
Bordetella pertussis 10 + 100 100 + 1000 100 + 1000<br />
Borrelia afzelii 100 + 320 + 1000 10<br />
Borrelia burgdorferi (strains CH and <strong>US</strong>A) 100 + 320 + 1000 10<br />
Borrelia garinii 100 + 320 + 1000 10<br />
Borrelia OspC 10<br />
Borrelia VlsE 100<br />
Campylobacter coli 100 100 + 1000 10<br />
Campylobacter jejuni 320 1000 100<br />
Candida albicans 1000 3200 + 10000 320<br />
Candida glabrata 1000 1000 + 3200 + 10000 320<br />
Candida krusei 1000 1000 + 3200 + 10000 320<br />
Candida parapsilosis 1000 1000 + 3200 + 10000 320<br />
Candida tropicalis 1000 1000 + 3200 + 10000 320<br />
Chikungunya virus 10 + 100 10 + 100<br />
Chlamydia pneumoniae (IFA) 100 100 + 1000 10<br />
Chlamydia pneumoniae (MIF) 10 100 10<br />
Chlamydia trachomatis (IFA) 100 320 + 1000 10<br />
Chlamydia trachomatis (MIF) 10 100 10<br />
Chlamydia psittaci (MIF) 10 100 10<br />
CMV 100 100 + 1000 100<br />
Coxsackie virus types A7., A9, A16, A24, B1 to B6 10 + 100 100 + 1000 10<br />
Crimean Congo fever virus 100 + 1000 10 + 100<br />
Dengue virus 100 + 1000 10 + 100<br />
EBV-CA, -EA 10 + 100 10 + 100 + 1000 10 + 100<br />
EBNA 10 + 100<br />
Echinococcus granulosus 100 100 + 320 + 1000 100<br />
Echo virus (type 7., 19) 10 + 100 100 + 1000 10<br />
Hanta virus 100 + 1000 100 + 1000<br />
Haemophilus influenzae 100 1000 + 10000 10<br />
Helicobacter pylori 32 + 100 100 + 1000 10<br />
HHV-6 10 + 100 10<br />
HSV-1/2 10 100 + 1000 + 10000 10<br />
Influenza virus type A 10 10 + 100 10<br />
Influenza virus type B 10 10 + 100 10<br />
Japanese encephalitis virus 10 + 100 + 1000 10 + 100<br />
Klebsiella pneumoniae 100 100 100<br />
Legionella pneumophila (all serotypes) 100 + 320 + 1000<br />
Leishmania 100 320 100<br />
Listeria monocytogenes (type 1/2a, 4b) 100 100 + 1000 100<br />
measles virus 10 + 100 10<br />
mumps virus 10 + 100 10<br />
Mycoplasma pneumoniae 10 10 + 100 10<br />
Parainfluenza virus types 1 - 4 10 + 100 10 + 100 + 1000 10<br />
Plasmodium falciparum/vivax 32 + 100<br />
Rift valley fever virus 100 + 1000 10 + 100<br />
rubella virus 10 + 100<br />
RSV 10 10 + 100 + 1000 10<br />
Saccharomyces cerevisiae 100 1000 100<br />
Sandfly fever virus 100 + 1000 100 + 1000<br />
SARS coronavirus 10 10 + 100 10<br />
TBE virus 10 10 + 100 + 1000 10 + 100<br />
Toxoplasma gondii 16+64+256 16+64+256+1028... 16+64+256<br />
Treponema pallidum 10 10 + 100 10<br />
VZV 10 10 + 100 10<br />
West-Nile virus 10 + 100 + 1000 10 + 100<br />
Yellow fever virus 100 + 1000 10 + 100<br />
Yersinia enterocolitica O:3; O:4; O:6; O:9 10 + 100 100 10 + 100<br />
— 15 —<br />
Investigation<br />
Techniques
<strong>EUROIMMUN</strong> Medizinische<br />
Labordiagnostika<br />
AG<br />
Investigation<br />
Techniques<br />
Prof. Dr. Winfried Stöcker<br />
Clinical Pathologist<br />
Ig- AGM A G M BASIS SPECTRUM<br />
151 ANA (cell nuclei) IF global testing<br />
152 ANA profile differentiation<br />
1572 dsDNA-NcX ELISA<br />
1572 dsDNA IFT SLE specific<br />
1590 ENA ProfilePlus ELISA 1<br />
nRNP/Sm, Sm, SS-A, SS-B, Scl-70, Jo-1<br />
1590 ENA ProfilePlus ELISA 2<br />
rib. P proteins, RNP/Sm, Sm, SS-A,<br />
SS-B, Scl-70, Jo-1, CENP B<br />
1590 SLE Profile ELISA (dsDNA, histones, rib.<br />
P prot., nRNP/Sm, Sm, SS-A, SS-B, Scl-70)<br />
162 AMA (mitochondria)<br />
171 ASMA (smooth muscle)<br />
120 cANCA* (granulocytes) Wegener’s dis.<br />
121 pANCA* (granulocytes) vasculitis<br />
1030 autoantibody profile 30 IF substrates<br />
Ig- AGM A G M ANA DIAGNOSTICS, WESTERNBLOT<br />
1520 PM-Scl, CENP A/B, Ku 86 and 72 kDa,<br />
M2 74 kDa, RNP 70 kDa, RNP A/C,<br />
Sm B/B’/D, SS-A 60 and 52 kDa, Ro-52,<br />
SS-B 52, 47, 44 and 43 kDa, ribosomal<br />
P proteins P0/P1/P2, Scl-70, Jo-1)<br />
Ig- AGM A G M OTHER AUTOANTIBODIES<br />
1612 centrioles<br />
1613 MSA-1 (NuMa, spindle fibres)<br />
1614 MSA-2 (midbody)<br />
1615 MSA-3 (chromosome-ass. antigen)<br />
1617 centromer F protein (CENP-F)<br />
1640 ribosomes<br />
1642 Golgi apparatus<br />
1643 lysosomes<br />
165 cytoskeleton<br />
1651 actin<br />
1652 vimentin<br />
1653 cytokeratin<br />
1654 tropomyosin<br />
1655 vinculin<br />
1656 desmin<br />
1659 laminin (basal membranes)<br />
1947 collagen type VII<br />
1950 elastin<br />
196 vessel endothelium<br />
Ig- AGM A G M THERAPY CONTROL<br />
1821 interferon alpha***<br />
1822 interferon beta<br />
1824 erythropoetin<br />
1572 dsDNA RIA<br />
1818 CIC-C1q ELISA<br />
— 16 —<br />
Clinical Immunology Laboratory<br />
Seekamp 31<br />
D-23560 Lübeck (Germany)<br />
Telephone +49 451 58 55 986<br />
Fax 58 55 134<br />
Diagnostically Relevant Autoantibodies<br />
Systemic Autoantibodies against<br />
Ig- AGM A G M SYST. LUP<strong>US</strong> ERYTHEMATOS<strong>US</strong> (SLE)<br />
151 ANA (cell nuclei) IF global testing<br />
1574 nucleosomes SLE specific<br />
1571 dsDNA ELISA SLE specific<br />
1572 dsDNA-NcX ELISA SLE specific<br />
1572 dsDNA IFT (C. luciliae) SLE specific<br />
1572 dsDNA RIA SLE specific<br />
159 ENA PoolPlus ELISA<br />
1591 U1-nRNP (70K, A, C)<br />
1593 Sm SLE specific<br />
1595 SS-A (Ro) 60 kDa: native<br />
159s Ro-52: recombinant<br />
1597 SS-B (La)<br />
1641 ribosomal P proteins SLE specific<br />
1605 Ku<br />
1601 cyclin I (PCNA)<br />
156 histones (global testing)<br />
1576 ssDNA (single-stranded DNA)<br />
121 pANCA* (granulocytes) vasculitis<br />
Ig- AGM A G M SJÖGREN’S SYNDROME<br />
151 ANA (cell nuclei) IF global testing<br />
1595 SS-A (Ro) 60 kDa: native<br />
159s Ro-52: recombinant<br />
1597 SS-B (La)<br />
Ig- AGM A G M ANTI-PHOSPHOLIPID<br />
SYNDROME (APS)<br />
1621 cardiolipin<br />
1632 ß-2-glycoprotein 1<br />
1631 lupus anticoagulant (plasma)**<br />
162a phosphatidylserine<br />
Ig- AGM A G M SYSTEMIC SCLEROSIS<br />
(DIFF<strong>US</strong>E + LIMITIERTE FORM)<br />
151 ANA (cell nuclei) IF global testing<br />
1532 Systemic Sclerosis Profile EUROLINE<br />
(Scl-70, CENP A, CENP B, RP11, RP155,<br />
Fibrillarin, NOR90, Th/To, PM-Scl100,<br />
PM-Scl75, Ku, PDGFR, Ro-52)<br />
1599 Scl-70 (DNA topoisomerase I)<br />
1584 PM-Scl75 (75 kDa)<br />
1584 PM-Scl100 (100 kDa)<br />
1611 centromeres<br />
1611 centromere A and B protein (rec.)<br />
1582 U3-nRNP (fibrillarin)<br />
1583 RNA polymerase I, II, III<br />
1585 7-2-RNP (Th/To)<br />
1586 4-6-S-RNA<br />
1587 NOR (nucleolus organizer region)<br />
1605 Ku<br />
CIRC. IMMUNE COMPLEXES<br />
1818 C1q ELISA<br />
Ig- AGM A G M SHARP’S SYNDROME MCTD<br />
1591 U1-nRNP (70K, A, C)<br />
151 ANA (cell nuclei) IF global testing<br />
Ig- AGM A G M POLYMYOSITIS, DERMATOMYOSITIS<br />
151 ANA (cell nuclei) IF global testing<br />
1530 Myositis Profile 3 EUROLINE<br />
(Mi-2, Ku, PM-Scl100, PM-Scl75, SRP, Jo-1,<br />
PL-7, PL-12, OJ, EJ, Ro-52)<br />
1661 Jo-1<br />
1662 PL-7<br />
1663 PL-12<br />
1664 OJ<br />
1665 EJ<br />
1584 PM-Scl75, PM-Scl100<br />
1616 SRP (signal recognition particle)<br />
159s Ro-52: recombinant<br />
1605 Ku<br />
1607 Mi-2<br />
1635 serotonin ab<br />
1636 PMR (polymyalgia rheumatica factor)<br />
Ig- AGM A G M (RHEUMATOID) ARTHRITIS<br />
1505 CCP (cyclic citrullinated peptides)<br />
151a Sa<br />
1814 RF (class. rheumatoid factor)<br />
1508 filaggrin (RA keratin)<br />
1219 GS ANA (granulocyte specific ANA)<br />
151 ANA (cell nuclei) IF global testing<br />
1604 RANA (rheum. arthritis nuclear antigen)<br />
121 pANCA* (granuloc.) RF-ass. vasculitis<br />
148 cartilaginous subst. polychondritis<br />
1947 collagen type VII<br />
FURTHER RHEUMATOID RELEVANT<br />
Ig- AGM A G M ANALYSES<br />
2011 anti-streptolysin<br />
2012 anti-streptokinase<br />
<strong>2013</strong> anti-streptodornase<br />
2014 anti-DPNase (anti-NADase)<br />
2031 anti-staphylolysin<br />
2034 anti-hyaluronidase<br />
213 Borrelia burgdorferi<br />
2171 Yersinia enterocolitica O:3<br />
2191 Chlamydia trachomatis<br />
IMMUNOGLOBULINS:<br />
Ig- AGM A G M ANTI-<br />
1811 human IgA<br />
1813 human IgE<br />
1814 human IgG<br />
1815 human IgM<br />
Grey boxes: standard analysis *) ANCA diagnostics in acute cases within one hour, at any hour **) Use special procedure for taking sample(s) ***) Send frozen sample(s)<br />
Fax:
<strong>EUROIMMUN</strong> Medizinische<br />
Labordiagnostika<br />
AG<br />
Prof. Dr. Winfried Stöcker<br />
Clinical Pathologist<br />
Ig- AGM A G M AUTOANTIBODY PROFILE<br />
1030 30 IF substrates (BIOCHIPs)<br />
Ig- AGM A G M THYROID GLAND<br />
1015 TRAb (TSH receptors)<br />
1012 TPO ab (thyroidea peroxidase)<br />
1013 TAb (thyroglobulin)<br />
1014 colloid antigen II ab<br />
1011 MAb (microsomes)<br />
1016 T 3 ab<br />
1017 T 4 ab<br />
Ig- AGM A G M DIABETES MELLIT<strong>US</strong><br />
1021 ICA (islet cell antibodies)<br />
1022 GAD (glutamic acid decarboxylase)<br />
1023 IA-2 (tyrosine phosphatase)<br />
1024 insulin ab human<br />
1025 insulin receptor<br />
1026 glucagon-producing cells<br />
1027 zink transporter 8<br />
147 lipocytes<br />
Ig- AGM A G M (POLY-)ENDOCRINOPATHY<br />
1051 adrenal cortex Addison’s dis.<br />
1053 21-hydroxylase Addison’s dis.<br />
1061 ovary: theca cells<br />
1062 ovary: corpus luteum<br />
1081 testis: Leydig cells<br />
105 steroid hormon-producing cells<br />
104 parathyroid gland<br />
1021 ICA (islet cell antibodies)<br />
1012 TPO ab (thyroidea peroxidase)<br />
1361 H + /K + -ATPase ab ELISA<br />
1361 PCA (parietal cells)<br />
1091 pituitary gland: anterior lobe<br />
1092 pituitary gland: posterior lobe<br />
1011 MAb (thyroid microsomes)<br />
1052 adrenal medulla<br />
107 placenta<br />
110 VPZ (vasopr.-prod. cells) D. insipudus<br />
Ig- AGM A G M INFERTILITY<br />
1621 cardiolipin<br />
1060 ovary: theca c., c. luteum, z. pellucida<br />
1081 testis: Leydig cells<br />
1086 spermatozoa<br />
1091 pituitary gland: anterior lobe<br />
107 placenta<br />
1401 prostate<br />
Ig- AGM A G M EPIDERMIS<br />
1501 desmosomes pemphigus<br />
1495 desmoglein 1 pemphigus<br />
1496 desmoglein 3 pemphigus<br />
1491 envoplakin paraneopl. pemphigus<br />
1502 epiderm. basal membr. pemphigoid<br />
1502 BP180 bullous pemphigoid<br />
1502 BP230 bullous pemphigoid<br />
135 oral mucosa Behçet’s/ Crohn’s dis.<br />
1503 basal membrane (urinary bladder)<br />
1509 epidermal keratin<br />
191 endomysium GSE, Duhring’s dis.<br />
3011 gliadin GSE, Duhring’s dis.<br />
1502 herpes gestationis factor<br />
1504 melanocytes<br />
150h hair follicle<br />
1947 collagen type VII NC1<br />
Ig- AGM A G M EYE<br />
1178 recoverin<br />
1177 tunica choroidea chron. chorioretinitis<br />
1171 cornea<br />
1172 retina<br />
1173 lens oculi<br />
1174 corpus ciliare<br />
1175 eye muscles<br />
1176 retro bulbar connective tissue<br />
120 cANCA* (granulocytes) Wegener’s dis.<br />
151 ANA (cell nuclei) IF global testing<br />
Ig- AGM A G M IMMUNOHAEMATOLOGY<br />
124 erythrocytes (global testing)<br />
1209 granulocyte membrane<br />
1221 lymphocytes<br />
1231 thrombocytes: indirect test (free ab)<br />
1232 thrombocytes: direct test (bound ab)**<br />
1361 H + /K + -ATPase ab ELISA<br />
1361 PCA (parietal cells)<br />
Clinical Immunology Laboratory<br />
Seekamp 31<br />
D-23560 Lübeck (Germany)<br />
Telephone +49 451 58 55 986<br />
Fax 58 55 134<br />
Diagnostically Relevant Autoantibodies<br />
Organ-/Tissue-Specifi c Autoimmunity: Autoantibodies against<br />
Ig- AGM A G M ANCA-ASSOC. VASCULITIDES<br />
(WEGENER’S DIS., MICR. ARTERITIS,<br />
CHURG-STRA<strong>US</strong>S SYNDROME)*<br />
120 cANCA IFT* granuloc. Wegener’s dis.<br />
1201 PR3 (proteinase 3)<br />
1202 BPI (CAP 57)<br />
121 pANCA IFT* granulocytes vasculitis<br />
1211 MPO (myeloperoxidase)<br />
1212 elastase<br />
1213 cathepsin G<br />
1215 lactoferrin<br />
120 ANCA Profile ELISA<br />
PR3, MPO, elastase, cath. G, BPI, lactoferrin<br />
151 ANA (cell nuclei) IF global testing<br />
195 elastin<br />
196 vessel endothelium<br />
Ig- AGM A G M KIDNEY, LUNG<br />
120 cANCA IFT* granuloc. Wegener’s dis.<br />
121 pANCA IFT* granulocytes vasculitis<br />
125 kidney IF global testing<br />
1251 GBM ELISA glomerular basal membrane<br />
1254 PLA2R idiop. membr. nephropathy<br />
151 ANA (cell nuclei) IF global testing<br />
1572 dsDNA IFT<br />
1252 TBM (tubular basal membrane)<br />
1271 lung alveolar basal membrane<br />
Ig- AGM A G M NERVO<strong>US</strong> SYSTEM<br />
111 Neuronal Ab IFT global testing<br />
Paraneoplastic neurol. syndromes<br />
1111 Neuronal Antigens Profile PL<strong>US</strong> RST<br />
EUROLINE amphiphysin, CV2, PNMA2<br />
(Ma-2), Ri, Yo, Hu, Recoverin, SOX1, Titin<br />
1112 Tr (Purkinje cell cytoplasm)<br />
1113 Yo (Purkinje cell cytoplasm; PCA-1)<br />
1114 PCA-2 (Purkinje cell cytoplasm)<br />
1115 Ri (neurone nuclei; ANNA-2)<br />
1116 Hu (neurone nuclei; ANNA-1)<br />
112d NMDA receptors<br />
112k AMPA receptors (GluR1, GluR2)<br />
112l GABA B receptors<br />
1117 Ma1/Ma2 (neurone nuclei; Ta)<br />
1119 CV2 (CRMP-5)<br />
1022 GAD stiff-person syndr.<br />
112e amphiphysin stiff-person syndr.<br />
112a AGNA (anti-glia nuclear antigen; SOX-1)<br />
112b ANNA-3<br />
1439 potassium channels (VGKC)<br />
1439 LGI1<br />
1439 CASPR2<br />
further parameters<br />
1154 aquaporin-4 neuromyelitis optica<br />
1157 glycine receptors<br />
1156 MOG (myelin-oligodendroc. glykoprot.)<br />
1121 myelin<br />
1122 MBP (myelin-basic protein)<br />
1123 MAG (myelin-assoc. glycoprotein)<br />
1124 myelin of peripheral nerves<br />
1126 neuroendothelium<br />
1127 neurofilaments<br />
1128 GFAP (glial fibrillary acidic protein)<br />
1129 non-medullated nerves<br />
112f astrocytes<br />
112g basal ganglia<br />
112h ganglion stellatum<br />
112i plexus myentericus<br />
1130 ganglioside profile<br />
GM 1 , GM 2 , GM 3 , GD 1a , GD 1b , GT 1b , GQ 1b<br />
113 single ganglioside analysis:<br />
GM 1 GM 2 GM 3 GD 1a<br />
GD 1b GT 1b GQ 1b<br />
151 ANA (cell nuclei) IF global testing<br />
213 Borrelia burgd.: serum CSF<br />
Ig- AGM A G M SKELETAL M<strong>US</strong>CLE, THYM<strong>US</strong><br />
1435 acetylcholine receptors M. gravis<br />
1434 MuSK M. gravis<br />
1437 calcium channels (VGCC) LEMS<br />
1439 potassium ch. (VGKC) neuromyotonia<br />
1439 CASPR2 neuromyotonia<br />
144 thymus M. gravis, thymoma<br />
1431 titin M. gravis<br />
143 skeletal muscle M. gravis<br />
1432 sarcolemma<br />
1436 myosin<br />
Ig- AGM A G M LIVER, BILIARY DUCTS<br />
130 Liver Ab IFT global testing, 6 BIOCHIPs<br />
130 Autoimmune Liver Dis. Ab Profile<br />
EUROLINE AMA-M2, 3E (BPO), Sp100,<br />
PML, gp210, LC-1, LKM-1, SLA/LP, Ro-52<br />
Autoimmune hepatitis (AIH)<br />
1302 SLA/LP (soluble liver antigen)<br />
1651 F-actin<br />
151 ANA (cell nuclei) IF global testing<br />
1307 LC-1 (liver cytosol)<br />
132 LKM (liver kidney microsomes)<br />
1321 LKM-1 ELISA<br />
1322 LKM-2<br />
1323 LKM-3<br />
1303 ASGPR (asialoglycoprotein receptors)<br />
171 ASMA (smooth muscles)<br />
1301 LSP (liver-specific protein)<br />
1304 LMA (liver cell membrane)<br />
Primary biliary cirrhosis (PBC)<br />
162 AMA (mitochondria)<br />
1622 AMA-M2 (PDH + BPO)<br />
1624 AMA-M4 (sulfitoxidase)<br />
1629 AMA-M9 (glycogen phosphorylase)<br />
1603 Sp100, PML (nuclear dots)<br />
1608 gp210 (nuclear membrane, lamin)<br />
Primary-sclerosing cholangitis (PSC)<br />
121 pANCA (granulocytes)<br />
further antibodies<br />
1305 bile ducts<br />
1306 bile canaliculi<br />
1609 coilin; P80 (few nuclear dots)<br />
Ig- AGM A G M STOMACH, INTESTINE<br />
1361 PCA (parietal cells) atroph. gastritis<br />
1361 H + /K + -ATPase ab atroph. gastritis<br />
1362 intrinsic factor ab vit. B 12 deficiency<br />
1366 gastrin (G) cells<br />
1391 pancreas acinus cells Crohn’s dis.<br />
1391 CUZD1 Crohn’s dis.<br />
1392 GP2 Crohn’s dis.<br />
2841 Saccharomyces cerev. Crohn’s dis.<br />
1392 pankreas secretion Crohn’s dis.<br />
135 mouth mucosa Behçet’s/ Crohn’s dis.<br />
1381 intestinal goblet cells ulc. colitis<br />
121 pANCA (granulocytes) ulc. colitis<br />
1382 enterocytes Crohn’s dis. , ulc. colitis<br />
191 endomysium GSE, Duhring’s dis.<br />
191 transglutaminase GSE, Duhring’s dis.<br />
3011 deamidated gliadin (Z-AGFA) GSE<br />
192 reticulin GSE, Duhring’s dis.<br />
EXOCRINE GLANDS, PANCREATITIS,<br />
Ig- AGM A G M SJÖGREN’S SYNDROME<br />
139 exocrine pancreas<br />
1391 pancreas acini<br />
1393 pancreas excretory ducts<br />
142 salivary glands (parotid gland)<br />
1421 parotid gland acini<br />
1423 parotid gland excretory ducts<br />
141 lacrimal gland<br />
Sjögren’s syndrome<br />
151 ANA (cell nuclei) IF global testing<br />
1595 SS-A (Ro)<br />
1597 SS-B (La)<br />
1576 ssDNA (single-stranded DNA)<br />
further antibodies<br />
1401 prostate<br />
1406 mamma<br />
Ig- AGM A G M HEART<br />
1627 AMA-M7 (myocard-specific)<br />
146 heart muscle<br />
1462 heart: intercalated disk<br />
1463 heart: myolemma<br />
Ig- AGM A G M LIPODYSTROPHY<br />
147 lipocytes<br />
Ig- AGM A G M ANTIBODIES AGAINST ANIMAL IgG<br />
3811 HAMA (human anti-mouse IgG) — 17. —<br />
Grey: standard *) ANCA diagn. in acute cases within 1 h **) Special procedure for taking sample ***) CV2 partial protein, which only contains the N-terminally localised epitopes of the antigen<br />
Investigation<br />
Techniques
<strong>EUROIMMUN</strong> Medizinische<br />
Labordiagnostika<br />
AG<br />
Investigation<br />
Techniques<br />
Prof. Dr. Winfried Stöcker<br />
Clinical Pathologist<br />
— 18 —<br />
IFT<br />
ELISA<br />
Westernblot<br />
EUROLINE<br />
Ig- AGM A G M BACTERIA A-Z<br />
219b Bartonella henselae<br />
219d Bartonella quintana<br />
2055 Bordetella parapertussis<br />
2050 Bordetella pertussis<br />
2131 Borrelia afzelii<br />
2132 Borrelia burgd. s. stricto<br />
CSF diagnostics<br />
2133 Borrelia burgd. (<strong>US</strong>A)<br />
2134 Borrelia garinii<br />
2092 Campylobacter coli<br />
2091 Campylobacter jejuni<br />
2192 Chlamydia pneumoniae<br />
2193 Chlamydia psittaci<br />
2191 Chlamydia trachomatis<br />
2040 Diphtheria toxoid<br />
2070 Haemophilus infl uenzae<br />
2080 Helicobacter pylori<br />
2101 Klebsiella pneumoniae<br />
2150 Legionella pneumophila<br />
serotypes ..................<br />
216 Legionella<br />
dumoffi i gormanii<br />
jordanis longbeachae<br />
micdadei<br />
2140 Listeria monocytogenes<br />
1/2a 4b<br />
2201 Mycoplasma hominis<br />
2202 Mycoplasma<br />
pneumoniae<br />
2060 Tetanus toxoid<br />
2111 Treponema pallidum<br />
CSF diagnostics<br />
2205 Ureaplasma urealyticum<br />
2170 Yersinia enterocolitica<br />
O:3 O:4 O:6 O:9<br />
Ig- AGM A G M PARASITES A-Z<br />
2320 Echinococcus gran.<br />
2231 Leishmania donovani<br />
2261 Plasmodium vivax<br />
2264 Plasmodium falciparum<br />
2410 Toxoplasma gondii<br />
Avidity determination<br />
Grey: standard analyses *) Currently not available as IVD in the European Union<br />
SI_0000_I_UK_A07, 07/2011<br />
Antibodies for Infectious Serology<br />
Respiratory tract<br />
Exanthema<br />
Lymphadenitis<br />
CNS<br />
Myocarditis<br />
Infectious arthritis<br />
Gastrointestinal tract<br />
Associated hepatitis<br />
Ophthalmology<br />
Antibodies against<br />
Otitis<br />
STD<br />
Pregnancy<br />
IFT<br />
ELISA<br />
Westernblot<br />
EUROLINE<br />
Fax:<br />
Clinical Immunology Laboratory<br />
Seekamp 31<br />
D-23560 Lübeck (Germany)<br />
Telephone +49 451 58 55 986<br />
Fax 58 55 134<br />
Ig- AGM A G M VIR<strong>US</strong>ES A-Z<br />
2680 Adenovirus type 3<br />
2730 Coxsackievirus type<br />
B1 B2 B3 B4 B5<br />
B6 A7 A9 A16 A24<br />
2570 Cytomegalovirus<br />
Avidity determination<br />
275a Echovirus type 7<br />
2791 Epstein-Barr virus<br />
capsid Ag (EBV-CA)<br />
Avidity determination<br />
2795 Epstein-Barr virus<br />
early Ag (EBV-EA)<br />
2793 Epstein-Barr virus<br />
nuclear Ag (EBNA)<br />
2531 HSV-1<br />
2532 HSV-2<br />
2511 HIV-1*<br />
2512 HIV-2*<br />
2536 Human herpes virus 6<br />
(HHV-6)<br />
2691 Infl uenza virus type A<br />
H1N1 H3N2<br />
2692 Infl uenza virus type B<br />
2610 Measles virus<br />
CSF diagnostics<br />
2630 Mumps virus<br />
CSF diagnostics<br />
2720 Parainfl uenza virus type<br />
1 2 3 4<br />
2670 Respiratory syncytial<br />
virus (RSV)<br />
2590 Rubella virus<br />
Avidity determination<br />
CSF diagnostics<br />
2661 TBE virus<br />
2650 Varicella zoster virus<br />
Avidity determination<br />
Ig- AGM A G M FUNGI A-Z<br />
2861 Candida albicans<br />
2862 Candida glabrata<br />
2863 Candida krusei<br />
2865 Candida parapsilosis<br />
2864 Candida tropicalis<br />
2841 Saccharom. cerevisiae<br />
Respiratory tract<br />
Exanthema<br />
Lymphadenitis<br />
CNS<br />
Myocarditis<br />
Infectious arthritis<br />
Gastrointestinal tract<br />
Associated hepatitis<br />
Ophthalmology<br />
Otitis<br />
STD<br />
Pregnancy
<strong>EUROIMMUN</strong> Medizinische<br />
Date of sample: Type of sample:<br />
Labordiagnostika<br />
AG<br />
Diagnosis:<br />
..................................................................................................................................................................................................<br />
GLOBAL TEST<br />
3840 Determination of total IgE (ELISA)<br />
INHALATION<br />
3110 Inhalation<br />
(g1, g3, g6, g12, t2, t3, t4, t7, w1, w6, w9,<br />
d1, d2, e1, e2, e3, m1, m2, m3, m6, CCD)<br />
3110 Inhalation 2<br />
(g6, g12, t2, t3, t4, w6, w9, d1, d2, e1, e2, e3, e6,<br />
e82, e84, es4, m1, m2, m3, m6, CCD)<br />
3111 Pediatric Inhalation<br />
(g6, g12, t2, t3, t4, w6, w8, w9, d1, d2, e1,<br />
e2, e3, e6, e82, e84, m1, m2, m3, m6, CCD)<br />
3112 Mediterranean Inhalation<br />
(g2, g6, t3, t4, t9, t11, t23, t210, w1, w6, w9, w19,<br />
d1, d2, d70, e1, e2, e3, m2, m6, CCD)<br />
3113 Inhalation „South East Asia“<br />
(ts19, t104, t19, t223, gs1, ds1, i6, u134, e1,<br />
e2, es172, e6, e71, e82, e84, ms1, ms4, m5,<br />
m12, m45, CCD)<br />
3116 Inhalation “China“<br />
(gs23, ts21, t3, t8, t11, t12, t14, t70, ws18, w1, w6,<br />
w9, es1, d1, d2, i6, ms5, m1, u73, u80, CCD)<br />
3116 Inhalation “China 2“<br />
(ds1, h1, i6, e1, e2, ms1, ts20, u80,<br />
w1, w6, CCD)<br />
3117 Inhalation “Middle East“<br />
(g1, g6, g12, t2, t3, t7, t9, w1, w6, w8, d1, d2,<br />
i6, e1, e84, m1, m2, m3, m5, m6, CCD)<br />
3118 Inhalation “Gulf“<br />
(g6, g12, t2, t3, t7, t9, w1, w6, d1, d2, i6, e1,<br />
e2, e3, e17, m1, m2, m3, m5, m6, CCD)<br />
3119 Inhalation “Turkey 1”<br />
(gs12, gs15, gs21, g12, ts23, ts24, t9, t70, ws18,<br />
ws19, ws20, d1, d2, i6, es2, es172, e1, e2, e3, e4,<br />
e80, e81, e84, ms11, ms12, m1, m2, m3, m6, CCD)<br />
3119 Inhalation “Top Screen”<br />
(ds1, e1, e2, e81, gs12, g12, t3, t9,<br />
w1, w6, w19, m2, IgE, CCD)<br />
3120 Inhalation “India“<br />
(g6, g12, g20, t18, w4, w27, w29, ds1, d2, i6, e1, e2,<br />
e11, e85, m3, m37, u81, u126, u129, u140, CCD)<br />
3121 Inhalation “Screen France“<br />
(hs12, es1, gs4, ts4, ws2, ms1, CCD)<br />
3210 SPAC Pollen 1<br />
(t3, g6, t215, t216, t220, t225, g205, g215,<br />
g210, g212, CCD)<br />
Serum .................................................................<br />
Antibodies for Allergology<br />
Sample number:<br />
Allergen Profi les: Antibodies of class IgE against<br />
FOOD<br />
3410 Food<br />
(f1, f75, f2, f45, f4, f5, f9, f13, f14,<br />
f17, f20, f49, f84, f237, f25, f31,<br />
f35, f85, f3, f23, CCD)<br />
3410 Food 2<br />
(f1, f75, f2, f78, f4, f5, f14, f10, f13,<br />
f17, f20, f49, f84, f95, f25, f31, f35,<br />
f85, f3, f23, CCD)<br />
3411 Food “South East Asia 1“<br />
(f1, f75, f2, f4, f9, f10, f14, f13, f17,<br />
f63, f64, f83, fs10, fs14, f23, f24,<br />
f80, f234, f105, f336, CCD)<br />
3411 Food “South East Asia 2“<br />
(f1, f75, f2, f4, f9, f10, f14, f13, f17, f63,<br />
f340, f83, fs10, fs14, f23, f24, f80, f234,<br />
f105, f336, CCD)<br />
3414 Food “China“<br />
(f1, f2, f4, f7, f27, f88, fs35, f13, f14,<br />
fs40, f25, f292, fs42, f23, f234, f3,<br />
f41, f56, fs41, fs77, CCD)<br />
3414 Food “China 2“<br />
(f1, f2, f13, f14, f23, f24, fs33, fs34, CCD)<br />
3415 Food “Middle East“<br />
(f1, f75, f2, f78, e204, f4, f14, f45, f13,<br />
f17, f20, f33, f49, f92, f25, f31, f85,<br />
f48, f88, f89, CCD)<br />
3416 Food “Gulf“<br />
(f1, f75, f2, f105, f4, f14, f45, fs36, f13,<br />
f92, f33, f44, f93, f25, f31, f48, f83,<br />
f88, f3, f23, CCD)<br />
3420 Food “Turkey 1”<br />
(f1, f75, f2, f169, f78, f4, f79, f9, f14, f10, f13, f17,<br />
f144, u87, f222, f73, f33, f44, f49, f92, f84, f146,<br />
f328, f25, f31, f35, f48, f95, f97, f122, f132, fs14,<br />
fs10, fs43, f83, CCD)<br />
3421 Food “India“<br />
(f2, f75, f168, f4, f9, f14, f13, f36, f49, f50,<br />
f35, f38, f48, f244, f83, f89, f74, f240,<br />
f23, f24, CCD)<br />
INSECT VENOMS<br />
3720 Insect Venoms<br />
(i1, i3, CCD)<br />
ATOPY, POLLEN-ASSOCIATED<br />
FOOD ALLERGIES<br />
3701 Atopy “Top Screen“<br />
(rs1, rs2, fx5, fs52, CCD)<br />
3710 Atopy<br />
(g6, g12, t3, w6, d1, e1, e2, e3, m2, m6, f1, f2,<br />
f3, f4, f9, f14, f17, f31, f35, f49, CCD)<br />
3710 Atopy 3<br />
(g6, t3, t4, w6, d1, d2, e1, e2, e3, m2, m3, f1,<br />
f75, f2, f3, f4, f13, f14, f31, f49, CCD)<br />
3711 Pollen-Food Cross Reactions<br />
(g6, t3, w6, f4, f5, f13, f17, f20, f48, f89,<br />
f271, f275, f44, f49, f348, f237, f328, f31,<br />
f35, f85, CCD)<br />
3712 Pediatrics<br />
(gx, t3, w6, d1, d2, e1, e2, e3, m2, m3, m6,<br />
f1, f75, f2, f3, f76, f77, f78, e204, f4, f9, f14,<br />
f13, f17, f31, f35, f49, CCD)<br />
3713 Atopy “China“<br />
(ts20, w1, w6, ds1, h1, e1, e2, i6, ms1, u80, f1, f2,<br />
f13, f14, f27, f88, fs33, fs34, f23, f234, CCD)<br />
3713 Atopy “China 3“<br />
(ts22, w6, us1, ds1, es1, h1, ms5, f245,<br />
fs9, fs43, fs56, fs34, fs44, CCD)<br />
3713 Atopy “China 4“<br />
(ds1, h1, i6, e1, e2, ms1, ts20, u80,<br />
w1, w6, f1, f2, f13, fs33, CCD)<br />
3716 Atopy “Peru“<br />
(e1, e2, e3, e7, e11, e82, e84, es172, i1, i3, i6,<br />
m1, m2, m3, m6, t18, u85, w29, w28, d1, d2,<br />
f75, f79, f1, f2, f23, f206, f13, f14, f104, f105,<br />
f26, fs32, f44, f49, CCD)<br />
EUROASSAY ALLERGY PROFILES<br />
3110 Inhalation<br />
(g1, g3, g6, g12, t2, t3, t4, t7, w1, w8, w6, w9, d1, d2,<br />
es4, e2, e3, e6, e1, e82, e84, m1, m2, m3, m6, CCD)<br />
3210 SPAC Pollen 1<br />
(t3, g6, t215, t216, t220, t225, g205, g215,<br />
g210, g212, CCD)<br />
3410 Food<br />
(f1, f75, f2, f78, f45, f4, f5, f9, f14, f10, f13, f17, f20,<br />
f84, f95, f237, f25, f31, f35, f85, f3, f23, f49, CCD)<br />
3712 Pediatrics/Atopy<br />
(g6, g12, t3, w6, d1, d2, e1, e2, e3, m2, m3, m6,<br />
f1, f75, f3, f2, f76, f77, f78, e204, f4, f9, f14, f13,<br />
f17, f31, f35, f49, CCD)<br />
Allercoat 6 System: Antibodies of class IgE against 600 different<br />
allergens of the areas animal allergens, environmental allergens,<br />
food, grasses, herbal and flower pollen, house dust, insects, mites,<br />
moulds, parasites, pharmaceutical drugs, trees.<br />
— 19 —<br />
Investigation<br />
Techniques
<strong>EUROIMMUN</strong> Medizinische<br />
Labordiagnostika<br />
AG<br />
Investigation<br />
Techniques<br />
POD-labelled<br />
anti-human antibody<br />
specific human<br />
antibody<br />
A<br />
B<br />
C<br />
D<br />
E<br />
F<br />
G<br />
H<br />
— 20 —<br />
substrate<br />
POD<br />
POD<br />
antigen-coated<br />
microplate well<br />
1 2 3 4 5 6 7 8 9 10 11 12<br />
<strong>EUROIMMUN</strong> Microplate ELISA<br />
stain<br />
chromogen<br />
Principle of the Test<br />
• Polystyrene microplate strips coated with purified, biochemically characterized<br />
antigens are used as solid phase containing bound antigens.<br />
• If the sample is positive, specific antibodies in the diluted serum sample attach<br />
to the antigens coupled to the solid phase.<br />
• In a second step, the attached antibodies are detected with peroxidase-labelled<br />
anti-human antibodies.<br />
• In a third step, the bound antibodies are made visible using a chromogen/<br />
substrate solution which is capable of promoting a color reaction. The intensity<br />
of the color produced is proportional to the concentration of antibodies in the<br />
serum sample.<br />
• Monospecific ELISA (enzyme immunoassays with a single antigen) provide a<br />
quantitative in vitro assay for the detection of antibodies.<br />
• “Profile ELISA” provide a semiquantitative in vitro assay for the detection of<br />
different antibodies on a single microplate strip.<br />
• The solid phase of “Pool ELISA” is coated with an antigen mixture for the semiquantitative<br />
detection of antibodies whose specificity must be subsequently<br />
investigated by monospecific assays.<br />
Reliable and Economical Calibration/Evaluation<br />
• In the case of a quantitative ELISA, calibration is generally performed using<br />
three cali bration sera.<br />
Calibration serum 1: upper limit of the measurement range<br />
Calibration serum 2: upper limit of the normal range (cut-off value)<br />
Calibration serum 3: negative<br />
• Only three wells are required for calibration, followed by serum samples. There<br />
is no need to incubate blank values or duplicate determinations.<br />
• Semiquantitative ELISA are performed using only one calibrator. Extended<br />
calibration curves are found in some ELISAs for immunity determination or CSF<br />
diagnostics.<br />
• The calibration is performed in relative units per milliliter (RU/ml) or, if an<br />
international reference serum exists, in international units per milliliter (IU/ml).<br />
• Each test can be optionally performed using a positive or negative control<br />
serum included in the test kit. Kit-specific reference ranges are provided for<br />
each calibrator and control serum.<br />
• All calibrations can be easily performed with the usual ELISA software.<br />
Easy, Quick and Economical Handling<br />
• Microplate strips containing break-off wells (except Profile ELISA).<br />
• To avoid mix-ups, microplate strips or reagent wells are printed with antigen<br />
abbreviations.<br />
• Reagents ready for use (wash buffer: concentrate).<br />
• Colour-coding allows clear identification of reagents.<br />
dark red: calibration serum 1 dark blue: pos. control serum<br />
red: calibration serum 2 green: neg. control serum<br />
light red: calibration serum 3 sample buffer and anti-human Ig PODconjugate:<br />
different colors<br />
• The sample buffer for infectious serology ELISA (detection of antibodies of class<br />
IgM) already contains an IgG/RF absorbent.<br />
• Compatible with all commercial washer and reader systems.
<strong>EUROIMMUN</strong> Medizinische<br />
Labordiagnostika<br />
AG<br />
Determination of Low-Avidity Antibodies<br />
<strong>EUROIMMUN</strong> Microplate ELISA<br />
• An alternative principle for the serological diagnosis of fresh infections has been<br />
established by investigating the antibody avidity.<br />
• The first reaction of the immune system following an infection is the formation<br />
of low-avidity antibodies. As the infection proceeds, increasingly antigen-adapted<br />
IgG is formed, and avidity grows. As long as high-avidity IgG is not yet detected<br />
in the serum, it can be assumed that the infection is still in an early stage.<br />
• To identify low-avidity antibodies in a patient’s serum, two microplate ELISA<br />
are performed in parallel: one test is carried out in the conventional way, the<br />
other one includes urea treatment between incubations with patient’s serum<br />
and per oxidase-labelled anti-human IgG, resulting in the detachment of lowavidity<br />
antibodies from the antigens.<br />
• Low-avidity antibodies are present if the ELISA extinction is significantly reduced<br />
by urea treatment. For an objective interpretation, the relative avidity index (RAI)<br />
can be calculated out of the measured values with and without urea incubation.<br />
• Serum dilution 1 : 101, conjugate class anti-human IgG, POD-labelled.<br />
• 3-point calibration, quantitative (IgG).<br />
• The following test kits for avidity determination are available: Toxoplasma gondii,<br />
CMV, rubella virus, measles virus, VZV, TBE virus, West-Nile virus, EBV-CA.<br />
Antibody Determination in CSF<br />
• Indication: local infections of the brain.<br />
• CSF dilution 1 : 2, serum dilution 1 : 404. Conjugate classes anti-human IgG or<br />
IgM, POD-labelled.<br />
• Easy to conduct: ready-for-use reagents.<br />
• 4-point calibration, quantitative. Identical incubation conditions and times (room<br />
temperature; 60 min / 60 min / 15 min): all <strong>EUROIMMUN</strong> ELISA for CSF diagnostics<br />
can be combined without difficulty on one and the same micro plate.<br />
• The antibody concentration in the patient’s serum is determined in parallel to<br />
the antibody concentration in CSF on one and the same microplate. The CSF/<br />
serum quotient CSQ is calculated from both measured values.<br />
path.-spec.<br />
• An intrathecal synthesis of specific antibodies is present if the CSF/serum<br />
quotient of the specific antibodies CSQ is significantly higher than the<br />
path.-spec.<br />
CSF/serum quotient of the whole IgG (CSQ ) or if necessary the CSQ .<br />
total lim.<br />
The relation of both values indicates the relative CSF/serum quotient CSQrel. (synonym: antibody specificity index, ASI).<br />
• Interpretation of results (according to the recommendations of Prof. Reiber):<br />
CSQ < 0,6: unreliable result, check for error<br />
rel.<br />
CSQ 0,6 – 1,3: standard range<br />
rel.<br />
CSQ 1,3 – 1,5: borderline range<br />
rel.<br />
CSQ > 1,5: Indication of pathogen-specific antibody production in the CNS<br />
rel.<br />
• For the automatic calculation of the CSQ <strong>EUROIMMUN</strong> provides a specific<br />
rel.<br />
Excel table free of charge.<br />
• Highest sensitivity, specificity and reproducibility. Antibody concentrations in<br />
serum and CSF can be determined over the total linear measurement range.<br />
• The following test kits for CSF diagnostics are available: Borrelia burgdorferi,<br />
Toxoplasma gondii, HSV-1, HSV-2, HSV-1/2 Pool, CMV, rubella virus, measles<br />
virus, mumps virus, VZV, TBE, EBV-CA.<br />
• All test systems for CSF diagnostics can also be used only for serology.<br />
• Perfectly adapted for the automated incubation in incubation devices.<br />
Number of Patients<br />
10<br />
8<br />
6<br />
4<br />
2<br />
RAI = E with urea<br />
E without urea<br />
0<br />
0 25 50 75 100<br />
Relative Avidity Index (RAI) in %<br />
Primary Infection<br />
Previous Infection<br />
Avidity of antibodies against EBV-CA (IgG)<br />
A CA<br />
B CB<br />
C CC<br />
D CD<br />
E<br />
F<br />
g<br />
h<br />
ELISA incubation scheme<br />
Table-based evaluation of the CSQ rel.<br />
CSQtotal<br />
1 2 3 4 5 6<br />
C1<br />
1:2<br />
S1<br />
1:404<br />
C2<br />
1:2<br />
S2<br />
1:404<br />
CA-D<br />
C3<br />
1:2<br />
S3<br />
1:404<br />
C4<br />
1:2<br />
S4<br />
1:404<br />
C5<br />
1:2<br />
S5<br />
1:404<br />
C6<br />
1:2<br />
S6<br />
1:404<br />
CSF-<br />
Calibrators C CSF S serum<br />
CSQalb.<br />
Reference range for normal values,<br />
intact blood-CSF barrier function<br />
Blood-CSF barrier dysfunction,<br />
no Ig production in CNS<br />
Blood-CSF barrier dysfunction,<br />
additional Ig production in CNS<br />
Clear Ig production in CNS,<br />
no disturbance in blood-CSF<br />
barrier function<br />
Error in blood extraction or analysis<br />
CFS/serum quotient diagram according to Reiber<br />
and Lange (1991)<br />
— 21 —<br />
Investigation<br />
Techniques
<strong>EUROIMMUN</strong> Medizinische<br />
Labordiagnostika<br />
AG<br />
Investigation<br />
Techniques<br />
— 22 —<br />
ELISA Automation Using the <strong>EUROIMMUN</strong> Analyzers<br />
approved<br />
approved<br />
<strong>EUROIMMUN</strong> Analyzer I and <strong>EUROIMMUN</strong> Analyzer I-2P: Broad<br />
Parameter Spectrum, Proven Reliability, Variable Throughput<br />
• One for all: fully automated processing of all <strong>EUROIMMUN</strong> ELISA – autoimmune<br />
diagnostics, infectious serology and allergology – with only one system<br />
• Flexibility for your benefit: open system with more than 800 validated<br />
•<br />
<strong>EUROIMMUN</strong> parameters for serum, plasma and cerebrospinal fluid, over 50 or<br />
30 parameters in parallel.<br />
Convenient, simple and reliable operation: e.g. by scanning QC certificates using<br />
a 2D-hand barcode scanner – ready-for-use reagents and preprogrammed assay<br />
protocols enable you to start immediately.<br />
• Capacity and throughput: quick loading and efficient time management allow<br />
processing of up to 70 or 50 tests per hour – up to 7 or 3 plates, 180 or 144<br />
samples at the start of a run.<br />
• Security for patients: validated test systems and the comprehensive safety kit<br />
provide the basis for reliable diagnostics.<br />
• Reliability and service: instruments and reagents from one manufacturer,<br />
quick and targeted support from our personnel for a smooth workflow in your<br />
laboratory.<br />
Modular System: A Highly Sophisticated Solution<br />
• High convenience, fast and reliable loading through barcode identification of<br />
samples and reagents: automatic scanning when racks are inserted, reading of<br />
lot-specific QC data via 2D hand barcode scanner.<br />
• Dilution area: 288 or 192 dilution positions (Deepwell, 2 ml).<br />
• Liquid level detection (capacitive), multi-shot (dispensing) mode, automatic tip<br />
detection, clot detection.<br />
• Pipetting possible during plate transport due to separation of transport and<br />
pipetting unit.<br />
• 4 or 2 incubators with heating and shaking function, 4 or 3 incubators at room<br />
temperature.<br />
• Standard Windows software: familiar user interface, all relevant statistical<br />
functions provided, available in different languages.<br />
• Efficient use and convenient handling of tips and dilution plates through memory<br />
function.<br />
A Convincing and Reliable Package: <strong>EUROIMMUN</strong> Analyzer,<br />
<strong>EUROIMMUN</strong> ELISA, <strong>EUROIMMUN</strong> Service<br />
• Comprehensive test system validation for the <strong>EUROIMMUN</strong> Analyzer: all<br />
para meters validated in accordance with the 98/7.9/EC directive and based on<br />
EN ISO 13485:2003/CMDCAS.<br />
• All ELISA test systems are manufactured according to the European Quality<br />
Standards (IVD).<br />
• National and international conformity (standardisation): CE, IVD, FDA and<br />
CMDCAS.<br />
• Programming and set-up of automated system, including introduction to the<br />
system with customer training, performed by qualified personnel.<br />
• Reliable and fast delivery of consumables and reagents.<br />
• Connection to in-house computer system via communication protocol ASTM.<br />
• Maintenance contract with <strong>EUROIMMUN</strong> on request.
<strong>EUROIMMUN</strong> Medizinische<br />
Labordiagnostika<br />
AG<br />
Pipette: 100 µl per well<br />
Incubate: 30 min<br />
Wash: 300 µl wash buffer<br />
per well<br />
Pipette: 100 µl per well<br />
Incubate: 30 min<br />
Wash: 300 µl wash buffer<br />
per well<br />
Pipette: 100 µl per well<br />
Incubate: 15 min<br />
Pipette: 100 µl per well<br />
Incubating the Microplate ELISA<br />
Evaluate: photometric measurement<br />
at a wavelength of 450 nm<br />
microplate wells coated<br />
with antigens<br />
diluted<br />
samples<br />
enzyme<br />
conjugate<br />
chromogen/<br />
substrate solution<br />
stop<br />
solution<br />
— 23 —<br />
Investigation<br />
Techniques
<strong>EUROIMMUN</strong> Medizinische<br />
Labordiagnostika<br />
AG<br />
Investigation<br />
Techniques<br />
chromogen<br />
nRNP/Sm<br />
— 24 —<br />
stain<br />
Sm<br />
SS-A<br />
SS-B<br />
membrane coated<br />
with antigen<br />
specific human<br />
antibody<br />
AP<br />
EUROASSAY: Line Blot in TITERPLANE Technique Format<br />
AP<br />
substrate<br />
AP-labelled<br />
anti-human<br />
antibody<br />
Scl-7.0<br />
Jo-1<br />
Control<br />
band<br />
EUROASSAY Anti-ENA ProfilePlus: detection of<br />
antibodies against SS-A and SS-B in a case of<br />
Sjögren’s syndrome.<br />
Principle of the Test<br />
• Membrane strips coated with thin parallel lines of several purified, biochemically<br />
characterized antigens are used as solid phase. The membrane strips are fixed<br />
as BIOCHIPs in the fields of microscope slides.<br />
• If the sample is positive, specific antibodies in the diluted serum sample attach<br />
to the antigens coupled to the solid phase.<br />
• In a second incubation step, the attached antibodies react with AP-labelled antihuman<br />
antibodies.<br />
• In a third step, the bound antibodies are stained with a chromogen/substrate<br />
solution which is capable of promoting a color reaction. An intense dark band<br />
at the line of the corresponding antigen appears if the serum sample contains<br />
specific antibodies.<br />
• The microscope slides are incubated using the TITERPLANE Technique:<br />
samples and reagents are applied to the reaction areas of a reagent tray. The<br />
slides are then placed into the recesses of the reagent tray, where all test strips<br />
of one slide come into contact with the liquids, and the individual reactions<br />
begin simultaneously.<br />
• Depending on the spectrum of antigens used, it is possible to analyze several<br />
antibodies next to each other and simultaneously under identical conditions.<br />
Easy Handling<br />
• Several serum samples can be analyzed simultaneously on one and the same<br />
slide.<br />
• Total time for performing the EUROASSAY test is about 100 minutes. During<br />
the washing procedure, reagents for the next incubation step can be applied to<br />
reagent trays.<br />
• All incubation steps proceed at room temperature. Shaking the slides together<br />
with the reagent tray on a circulatory shaker ensures the best possible<br />
sensitivity.<br />
• Low reagent consumption. Only 50 µl each of diluted serum and reagent are<br />
needed per test field.<br />
• Reagents ready for use (wash buffer: concentrate).<br />
Reliable and Simple Evaluation<br />
• Since results are evaluated visually, there is no investment required for<br />
photometers, etc.<br />
• The antigen bands are located at exactly defined positions, which means that<br />
evaluation of the test is much simpler than for Westernblots.<br />
• Correct completion of the individual incubation steps in each test field is<br />
indicated by staining of the control band.<br />
• Positive and negative results can be easily and reliably differentiated from each<br />
other. The intensity of the antigen bands correlates with the antibody titer.<br />
• The antigens used are highly pure, mostly isolated by affinity chromatography.<br />
The membrane strips do not contain any superfluous proteins which might<br />
cause unspecific positive results.<br />
• The incubated microscope slides can be stored for long periods. Results can be<br />
easily documented.
<strong>EUROIMMUN</strong> Medizinische<br />
Labordiagnostika<br />
AG<br />
Incubating the EUROASSAY (TITERPLANE Technique)<br />
Pipette: 50 µl per field<br />
Incubate: 30 min shaking on a<br />
circulatory shaker<br />
(300 rpm)<br />
Wash: 5 s flush<br />
15 min cuvette<br />
Pipette: 50 µl per field<br />
Incubate: 30 min shaking on a<br />
circulatory shaker<br />
(300 rpm)<br />
Wash: 5 s flush<br />
15 min cuvette<br />
Pipette: 50 µl per field<br />
Incubate: 10 min shaking on a<br />
circulatory shaker<br />
(300 rpm)<br />
Wash: flush with deionized<br />
or distilled water,<br />
air dry<br />
Evaluate: visual assessment<br />
of color intensity<br />
slide<br />
membrane strips<br />
wash<br />
buffer<br />
wash<br />
buffer<br />
reagent tray<br />
diluted samples<br />
enzyme conjugate<br />
chromogen/substrate<br />
solution<br />
— 25 —<br />
Investigation<br />
Techniques
<strong>EUROIMMUN</strong> Medizinische<br />
Labordiagnostika<br />
AG<br />
Investigation<br />
Techniques<br />
— 26 —<br />
The EUROLINE: A New Technique for Extensive Antibody Profiles<br />
AP-labelled<br />
anti-human<br />
antibody<br />
specific human<br />
antibody<br />
AMA M2<br />
M2-3E<br />
(BPO)<br />
Sp100<br />
PML<br />
gp210<br />
LKM-1<br />
LC-1<br />
SLA/LP<br />
Ro-52<br />
Control<br />
nRNP/Sm<br />
Sm<br />
SS-A<br />
Ro-52<br />
SS-B<br />
Scl-7.0<br />
PM-Scl<br />
Jo-1<br />
CENP B<br />
PCNA<br />
dsDNS<br />
Nucleos.<br />
Histones<br />
Rib. P-prot.<br />
AMA-M2<br />
substrate<br />
AP<br />
membrane coated<br />
with antigen<br />
Control<br />
stain<br />
chromogen<br />
AP<br />
Mi-2<br />
Ku<br />
PM-Scl100<br />
PM-Scl7.5<br />
Jo-1<br />
SRP<br />
PL-7.<br />
PL-12<br />
EJ<br />
OJ<br />
Ro-52<br />
Control<br />
Incubated EUROLINE test strips (Autoimmune<br />
Liver Diseases Profile, ANA Profile 3, Myositis<br />
Profile 3.<br />
Principle of the Test<br />
• Membrane strips coated with thin parallel lines of several purified, biochemically<br />
characterized antigens are used as solid phase. The membranes are fixed as<br />
BIOCHIPs onto synthetic foil.<br />
• If the sample is positive, specific antibodies in the diluted serum sample attach<br />
to the antigens coupled to the solid phase.<br />
• In a second incubation step, the attached antibodies react with alkalinephosphatase-labelled<br />
anti-human antibodies.<br />
• In a third step, the bound antibodies are stained with a chromogen/substrate<br />
solution which is capable of promoting a color reaction. An intense dark band<br />
at the line of the corresponding antigen appears if the serum sample contains<br />
specific antibodies.<br />
• Depending on the spectrum of antigens used, it is possible to analyze several<br />
antibodies next to each other and simultaneously under identical conditions.<br />
Easy Handling, Reliable and Simple Evaluation<br />
• A separate membrane strip is incubated for each serum sample.<br />
• Total time for performing the analysis is about 105 minutes.<br />
• The incubation can be automated using the EUROBlotMaster.<br />
• All incubation steps proceed at room temperature.<br />
• The antigen bands are located at exactly defined positions, which means that<br />
the evaluation of the test is much simpler than for Westernblots.<br />
• Correct completion of the individual incubation steps is indicated by staining of<br />
the control band on each EUROLINE test strip.<br />
• Positive and negative results can be easily and reliable differentiated from each<br />
other. The intensity of the antigen bands correlates with the antibody titer.<br />
• The antigens used are highly pure, mostly isolated by affinity chomatography.<br />
The membrane strips do not contain any superfluous proteins which might<br />
cause unspecific positive results.<br />
• The incubated EUROLINE test strips can be stored for long periods. Results can<br />
be easily documented.<br />
• The program EUROLineScan from <strong>EUROIMMUN</strong> has been developed to enable<br />
quantitative evaluation of EUROLINE test strips, to facilitate management of<br />
data, and to provide detailed documentation of results. First, the incubated<br />
EUROLINE test strips are scanned using a flatbed scanner. EUROLineScan<br />
recognizes the position of the strips, even if they have been placed inexactly,<br />
identifies the bands, and measures their intensity. The results are then saved<br />
together with the image data. A separate results sheet can be produced for each<br />
patient.
<strong>EUROIMMUN</strong> Medizinische<br />
Labordiagnostika<br />
AG<br />
EUROBlotMaster<br />
Modern Automation of Immunoblot Assays<br />
• Standardisation of immunoblot strips – higher precision and reproducibility.<br />
• Automatisation of all <strong>EUROIMMUN</strong> immunoblot strips (EUROLINE, EUROLINE-<br />
WB, Westernblot).<br />
• Over 90 validated profiles available (autoantibody diagnostics, infectious<br />
serology and allergology).<br />
• Two models available: for up to 30 or 44 strips per test run.<br />
• Easy operation.<br />
• Combination of different conjugates/tests in one run.<br />
• Walk-away function – fully automated from the start to the end of processing<br />
following loading.<br />
• Compatible with modern evaluation systems such as EUROBlotCamera and<br />
EUROLineScan.<br />
EUROBlotOne: Unique Automation Solution for the Immunoblot<br />
Work Station: From Sample Identification to Final Results<br />
• Fully automated system: Processing of immunoblot tests – from sample<br />
identification to the final result.<br />
• Security: Integreted barcode identification ensures that the correct samples are<br />
pipetted.<br />
• Flexibility: Autoimmune diagnostics, infectious serology and allergology on one<br />
system; many innovative multiparameter tests available.<br />
• High walk-away capacity: Up to 44 samples can be analysed in one run.<br />
• Simplified routine diagnostics: User-friendly software and hardware; minimal<br />
maintenance requirements.<br />
• Reliability: Automated evaluation using the worldwide established and customerfriendly<br />
EUROLineScan software; bidirectional connection to LIS available (via<br />
EUROLineScan).<br />
• All-in-one: One supplier providing complete service and support for reagents,<br />
automated processor and software.<br />
Automated Evaluation of Results Using EUROLineScan<br />
• For all <strong>EUROIMMUN</strong> blot systems: EUROLINE, EUROLINE-WB, Westernblot.<br />
• For all areas: autoimmune diagnostics, infectious serology and allergology.<br />
• EUROBlotCamera: digitalisation of strips while in the incubation tray.<br />
• EUROBlotScanner: digitalisation of strips using flatbed scanner.<br />
• Fully automated identification, quantitation and assignment of bands.<br />
• Option to modify results (changes are automatically documented).<br />
• Complete results obtained just a few minutes after finishing the incubation.<br />
• Fully automated administration and documentation of extensive individual data.<br />
• Electronic archiving of all images and data (avoids the need to store potentially<br />
infectious blot strips).<br />
• Online connection to laboratory software.<br />
• Network-compatible.<br />
EUROBlotMaster<br />
EUROBlotOne<br />
EUROLineScan Evaluation<br />
— 27. —<br />
Investigation<br />
Techniques
<strong>EUROIMMUN</strong> Medizinische<br />
Labordiagnostika<br />
AG<br />
Investigation<br />
Techniques<br />
Westernblot/EUROLINEWB: Reliable Differentiation of Antibodies Present<br />
— 28 —<br />
AP-labelled<br />
anti-human<br />
antibody<br />
specific human<br />
antibody<br />
EUROLINE-WB Borrelia<br />
substrate<br />
AP<br />
p 83<br />
p 41, Flag.<br />
p 39, Bmp A<br />
p 31, Osp A<br />
p 30<br />
p 25, Osp C<br />
p 21<br />
p 19<br />
p 17.<br />
AP<br />
membrane coated<br />
with antigen<br />
stain<br />
Alignment bar<br />
Control band<br />
chromogen<br />
VlsE antigen<br />
on EURO LINE<br />
membrane chip<br />
EUROLINE-WB: detection of antibodies against<br />
Borrelia.<br />
Principle of the Test<br />
• Membrane strips containing electrophoretically separated antigen extracts are<br />
used as solid phase. The position of the proteins depends on their respective<br />
molecular masses.<br />
• If the sample is positive, specific antibodies in the diluted serum sample attach<br />
to the antigens coupled to the membrane.<br />
• In a second incubation step, the attached antibodies react with AP-labelled antihuman<br />
antibodies.<br />
• In a third step, the bound antibodies are stained with a chromogen/substrate<br />
solution which is capable of promoting a color reaction. An intense dark band<br />
at the line of the corresponding antigen appears if the serum sample contains<br />
specific antibodies.<br />
• Evaluating the band patterns on the incubated membrane strips involves<br />
differentiating non-specific from specific antibodies. The number and intensity<br />
of the specific bands is decisive for the result “positive/negative“.<br />
Easy Handling, Reliable Evaluation and High Diagnostic Significance<br />
• A separate membrane strip is incubated for each serum sample.<br />
• Total time for performing the Westernblot test is about 115 minutes.<br />
• All incubation steps proceed at room temperature.<br />
• The bands are assigned according to a lot-specific evaluation matrix provided.<br />
A separate lot is issued for each electropho resis gel, helping to avoid errors in<br />
the assignment of the bands.<br />
• Every test kit contains a membrane strip of the same lot incubated with a<br />
positive reference serum. Therefore, there is no need to incubate a positive<br />
control serum.<br />
• The membrane strips are pre-numbered to prevent confusion. Laborious<br />
labelling is not necessary.<br />
• Correct completion of the individual incubation steps for each membrane strip<br />
is indicated by staining of the control band at the bottom of the strip.<br />
• Positive and negative reactions can be easily and reliably differentiated from<br />
each other. The intensity of the antigen bands correlates with the antibody titer.<br />
• The Westernblot is the method of choice when the objective is to confirm or<br />
differentiate positive results obtained in a screening test (indirect immunofluorescence<br />
or microplate ELISA).<br />
• EUROLINE-WB is a combination of westernblot and line blot techniques. Proteins<br />
from a whole antigen extract are electrophoretically separated according to<br />
molecular mass and transferred onto a nitrocellulose membrane. Highly purified<br />
native or recombinant antigens are then printed as lines onto the westernblot<br />
strips (EUROLINE membrane chip).<br />
• The program EUROLineScan from <strong>EUROIMMUN</strong> has been developed to enable<br />
quantitative evaluation of Westernblot/EUROLINE-WB test strips, to facilitate<br />
management of data, and to provide detailed documentation of results. First,<br />
the incubated Westernblot/EUROLINE-WB test strips are scanned using a flatbed<br />
scanner. EUROLineScan recognizes the position of the strips, even if they have<br />
been placed inexactly, identifies the bands, and measures their intensity. The<br />
results are then saved together with the image data. A separate results sheet<br />
can be produced for each patient.
<strong>EUROIMMUN</strong> Medizinische<br />
Labordiagnostika<br />
AG<br />
Incubating the EUROLINE/Westernblot/EUROLINEWB<br />
using the EUROBlotMaster or manually on a rocking platform<br />
Pipette: 1.5 ml per channel<br />
Incubate: 5-15 min shaking, depending<br />
on test system<br />
Aspirate off:<br />
Pipette: 1.5 ml per channel<br />
Incubate: 30 min shaking<br />
Wash: 1.5 ml buffer,<br />
5 min incubation,<br />
aspirate off<br />
Pipette: 1.5 ml per channel<br />
Incubate: 30 min shaking<br />
Wash: 1.5 ml buffer,<br />
5 min incubation,<br />
aspirate off<br />
Pipette: 1.5 ml per channel<br />
Incubate: 10 min shaking<br />
Wash: rinse with distilled<br />
water, aspirate off<br />
Evaluate: with EUROLineScan or<br />
visual assessment<br />
membrane strip<br />
incubation channel<br />
buffer<br />
diluted samples<br />
buffer<br />
enzyme conjugate<br />
buffer<br />
chromogen/substrate solution<br />
Applicable for most EUROLINE/Westernblot/EUROLINE-WB test kits<br />
— 29 —<br />
Investigation<br />
Techniques
<strong>EUROIMMUN</strong> Medizinische<br />
Labordiagnostika<br />
AG<br />
Investigation<br />
Techniques<br />
— 30 —<br />
The EUROArray: DNA Microarray Test Systems for Diagnostics (IvD)<br />
Blood sample DNA isolation<br />
Patient DNA<br />
DNA contains target sequence<br />
Primer<br />
1 st PCR cycle<br />
2 nd PCR cycle<br />
Next cycles<br />
Exponential DNA amplification<br />
and fluorescence labelling<br />
Binding of PCR products to<br />
complementary DNA probes<br />
of the microarray<br />
Carrier material DNA spots<br />
DNA does not contain target sequence<br />
Primer<br />
1 st PCR cycle<br />
2 nd PCR cycle<br />
Next cycles<br />
No DNA amplification<br />
No binding without<br />
complementary PCR products<br />
Carrier material DNA spots<br />
EUROArray slide and BIOCHIP after incubation.<br />
Principle of the Test<br />
Sample Preparation<br />
• DNA isolation: In order to investigate with a microarray if a patient‘s DNA<br />
contains particular sequences, the DNA must first be extracted from the patient‘s<br />
blood. This is performed, for example, using DNA isolation kits.<br />
Amplification of Patient DNA: Polymerase Chain Reaction (PCR)<br />
• The sections of DNA to be investigated are amplified million-fold using the<br />
polymerase chain reaction (PCR).<br />
• Two starter DNA molecules (primers) define the region to be copied. If the<br />
patient DNA contains the corresponding section (target sequence), the primers<br />
bind and the target sequence is copied.<br />
• This reaction is repeated many times, so that the DNA region between the<br />
primers is greatly (exponentially) amplified.<br />
• The resulting PCR products are labelled with a fluorescent dye, which enables<br />
them to be detected subsequently by the microarray. If the target sequence is<br />
not present in the patient sample, then the primers cannot bind and the DNA is<br />
not amplified.<br />
Analysis of PCR <strong>Product</strong>s on the Microarray: DNA Microarray Hybridisation<br />
• The PCR products are incubated with the microarray.<br />
• They are fi rst mixed with a hybridisation buffer, which provides optimal<br />
conditions for binding of the PCR products to the complementary probes on<br />
the microarray.<br />
• This binding is measured via the fl uorescence signals emitted by the spots.<br />
Advantages of the EUROArray System<br />
• Array platform based on proven BIOCHIP Technology.<br />
• Ready-to-use PCR components.<br />
• Simple procedure.<br />
• Standardised incubation using established TITERPLANE Technique.<br />
• Integrated control reactions secure sensitivity and specificity for every test<br />
sample.<br />
• Fully automated standardised evaluation, interpretation and archiving of results<br />
(EUROArrayScan software).<br />
• No DNA isolation with EUROArray Direct: the analysis is performed directly<br />
from pretreated blood.<br />
• Complete process from receipt of samples to issuing of results is IVD validated<br />
and CE labelled (DNA extraction, test kits, microarray scanner, software).<br />
• Pre-prepared LIMS connection.
<strong>EUROIMMUN</strong> Medizinische<br />
Labordiagnostika<br />
AG<br />
Performance and Evaluation of the EUROArray Test<br />
Save Time and Money with the “EUROArray Direct“ Procedure<br />
• For some parameters <strong>EUROIMMUN</strong> offers a rapid procedure in which DNA<br />
isolation is not necessary.<br />
• The blood sample is incubated with extraction solution provided in the kit (E 1<br />
and E 2, see figure). The DNA extract is used directly in the PCR.<br />
Comparison EUROArray EUROArray Direct<br />
Blood volume ~ 200 µl 5 µl<br />
Time DNA isolation: DNA extraction:<br />
approx. 80 min for 40 samples < 20 min. for 40 samples<br />
Costs For DNA isolation kit No additional costs, extraction<br />
solutions contained in the kit<br />
Simple, uncomplicated and effortless<br />
• All PCR reagents supplied in EUROArray kits are ready for use, including the<br />
DNA polymerase and the validated specific primers. Thus, the number of<br />
pipetting steps is reduced to a minimum and laborious optimisation processes<br />
are eliminated.<br />
• The PCR works reliably with minimal effort: the pre-prepared PCR reagents are<br />
simply combined, and the DNA is then added to this master mix.<br />
• The DNA microarray hybridisation is performed under exact, standardised<br />
conditions using the proven TITERPLANE Technique. This procedure is simple<br />
and reliable. The samples (PCR products + hybridisation buffer) are pipetted<br />
onto the reaction fields of a reagent tray. The slides are then placed into the<br />
recesses of the reagent tray, whereby all BIOCHIPs come into contact with the<br />
liquids simultaneously. Thanks to the hydrophobic surroundings, the fluid drops<br />
remain stable on the hydrophilic reaction fields during the incubation and do not<br />
run into one another.<br />
• After a one-hour incubation period in the hybridisation station, the EUROArray<br />
slides are washed: 10 slides are processed in just 5 minutes and can then be<br />
evaluated.<br />
Fully automated standardised evaluation delivers fast and reliable<br />
results<br />
• With the <strong>EUROIMMUN</strong> Microarray Scanner and EUROArrayScan software,<br />
EUROArrays are evaluated easily, quickly and objectively without the need to<br />
study complicated manuals.<br />
• EUROArrayScan software can be integrated into EUROLabOffice and other<br />
laboratory information management systems (LIMS) without any difficulties.<br />
• At the start of each run, the data for the samples to be examined are entered<br />
and are then transferred automatically into the working list by the software.<br />
• After the incubated slides have been placed into the microarray scanner, the<br />
scanning procedure is initiated simply by a mouse click.<br />
• EUROArrayScan software evaluates all data fully automatically, produces a<br />
report and documents and archives all results.<br />
• Results for a EUROArray slide (up to five samples) are obtained in less than 20<br />
seconds!<br />
5 µl Blood<br />
To be used<br />
directly for PCR<br />
Pipette samples<br />
Reagent tray<br />
Incubate<br />
20 µl E1<br />
5 sec<br />
1 min<br />
20 µl E2<br />
DNA extraction with the „Direct procedure”.<br />
+<br />
Ready for use<br />
PCR reagents PCR master mix<br />
PCR<br />
Place slide onto tray<br />
1 hour<br />
Hybridisation station with four<br />
EUROArray slides<br />
— 31 —<br />
Investigation<br />
Techniques
<strong>EUROIMMUN</strong> Medizinische<br />
Labordiagnostika<br />
AG<br />
Investigation<br />
Techniques<br />
125 I<br />
— 32 —<br />
radioactively<br />
labelled<br />
antigen<br />
(“tracer”)<br />
solid phase<br />
(tube surface)<br />
<strong>EUROIMMUN</strong> Radioimmunoassays (RIA/IRMA)<br />
precipitation<br />
agent<br />
antibody<br />
specific human antibody<br />
radioactively<br />
labelled<br />
antibody<br />
analyte<br />
125 I<br />
125 I<br />
Test principle RIA (precipitation techniques)<br />
• In the first incubation step patient sera are incubated with 125 I-labelled antigen in<br />
polystyrene tubes. Any specific antibodies in the sera bind to the antigen.<br />
• In the second incubation step the antigen-antibody complexes are precipitated<br />
using a precipitation agent.<br />
• The precipitate is washed with buffer. After centrifugation and decanting of<br />
the supernatant, the radioactivity in the precipitate is counted using a gamma<br />
counter. The intensity of the radioactivity is proportional to the concentration of<br />
specific antibody in the patient serum.<br />
• The antibody concentration is evaluated quantitatively using a calibration curve.<br />
Test principle RIA (coated tubes)<br />
• RIA tests (coated tubes) are competitive ligand assays for antibody and antigen<br />
determinations.<br />
• The intensity of radioactive radiation is inversely proportional to the concentration<br />
of specific antibodies or antigens in the sample.<br />
• The quantitative evaluation of the antigen/antibody concentration is carried out<br />
using a calibration curve.<br />
Test principle IRMA (antigen determination)<br />
• With this test principle, monoclonal antibodies which are bound directly or<br />
indirectly to the inner wall of polystyrene tubes are used.<br />
• During the first incubation step, the patient sera to be investigated are incubated<br />
with the monoclonal 125I-labelled antibodies in the coated tubes.<br />
• The antigen in the sample is bound by the immobilised antibodies and by the<br />
125I-labelled antibodies. This results in a solid-phase bound sandwich complex.<br />
• The unbound 125I-labelled antibodies are removed by washing and subsequently<br />
decanting. The intensity of radioactive radiation is proportional to the concentration<br />
of antigens in the patient serum.<br />
• The quantitative evaluation of the antigen concentration is carried out using a<br />
calibration curve.<br />
Simple and economical handling, reliable analysis<br />
• Simple test procedure.<br />
• Synchronous processing of samples, including different tests in parallel.<br />
• Ready-to-use reagents (possible exceptions: tracer, precipitation reagents).<br />
• Different test formats for small and large sample series.<br />
• Because of the large measurement range of <strong>EUROIMMUN</strong> RIA, further<br />
•<br />
measurements with other sample dilutions are generally not necessary.<br />
Simple evaluation of test results.<br />
• Each test can be optionally evaluated with controls which are supplied in the<br />
test kit. Test kit-specific reference ranges are given for all controls.<br />
• <strong>EUROIMMUN</strong> radioimmunoassays show the following analytical characteristics:<br />
– High analytical specificity.<br />
– High detection sensitivity.<br />
– High clinical sensitivity and specificity.<br />
– Good reproducibility.
<strong>EUROIMMUN</strong> Medizinische<br />
Labordiagnostika<br />
AG<br />
<strong>EUROIMMUN</strong> PRODUCTS FOR ThE DETERMINATION OF AUTOANTIBODIES<br />
— 33 —<br />
<strong>EUROIMMUN</strong> <strong>Product</strong>s for the<br />
Determination of Autoantibodies
<strong>EUROIMMUN</strong> Medizinische<br />
Labordiagnostika<br />
AG<br />
<strong>EUROIMMUN</strong> <strong>Product</strong>s for the<br />
Determination of Autoantibodies<br />
EUROPL<strong>US</strong> ANA Mosaic 20A (HEp-20-10 cells,<br />
pri mate liver, SS-A+SS-B BIOCHIPs, rib. P-prot.+<br />
Jo-1 BIOCHIPs: antibodies against SS-A/SS-B.<br />
Order No. Formats<br />
FA 1512-####-20 page 131<br />
HEp-2010: antibodies against spindle fibers.<br />
Order No. Formats<br />
FA 1522-#### page 131<br />
Incubated EUROASSAY Anti-ENA ProfilePlus.<br />
Order No. Formats<br />
DA 1590-####-1 G page 80<br />
— 34 —<br />
Autoantibodies against Cell Nuclei (ANA)<br />
Indirect Immunofluorescence Test: EUROPL<strong>US</strong> ANA Mosaic 20A<br />
• Screening test for the detection of antibodies against cell nuclei (ANA).<br />
• Indications: rheumatic diseases.<br />
• Initial dilution 1 : 100; conjugate class anti-human IgG, FITC-labelled.<br />
• Using HEp-20-10 cells many antibodies against cell nuclei can be analyzed, e.g.<br />
anti bodies against DNA, histones, RNA, nRNP, Sm, SS-A, SS-B, nuclear dots,<br />
centro meres, nuclear membrane, nucleoli (PM-Scl, fibrillarin, RNA polymerase<br />
I, NOR), Scl-7.0, cyclin I and II, spindle fibers, midbody, centrioles.<br />
• In addition, cytoplasmic autoantibodies are identified with HEp-20-10 cells: antibodies<br />
against ribosomes, Jo-1, mito chondria, Golgi apparatus, actin etc.<br />
• The primate liver permits the verification of results between both substrates, makes<br />
the pre-differentiation of a large number of ANA possible, and helps to establish<br />
titer levels. Moreover, the primate liver often contains additional antigens,<br />
allowing the identification of further autoantibodies: antibodies against LMA, LSP,<br />
endo mysium, bile ducts and endothelium cells, as well as cANCA und pANCA.<br />
• The EUROPL<strong>US</strong> system is a monospecific test that can be used to confirm the<br />
presence of autoantibodies against cell nuclei and cytoplasm with one and the<br />
same test kit. The following antigens are currently available as EUROPL<strong>US</strong><br />
BIOCHIPs: SS-A, SS-B, nRNP/Sm, Sm, Scl-7.0, Jo-1, ribosomal P-proteins.<br />
Indirect Immunfluorescence Test: Innovative Cell Line HEp-20-10<br />
with a high number of mitotic cells<br />
• Screening test for detection of antibodies against cell nuclei.<br />
• Indications: rheumatic diseases.<br />
• Initial dilution 1 : 100; conjugate class anti-human IgG, FITC-labelled.<br />
• Compared to standard HEp-2 cells, HEp-20-10 cells demonstrate 10-fold more<br />
mitotic cells. Antibodies against mitotic-specific structures (centromeres, spindle<br />
fibers, midbody, centrioles, NOR) can be more easily identified than with conventional<br />
preparations.<br />
• At the same time, the cell line HEp-20-10 offers the full antigen spectrum for cell<br />
nuclei antibody diagnostics.<br />
• The BIOCHIP with HEp-20-10 can be supplemented with additional substrates,<br />
for example, primate liver (ANA; Order No. FA 1512-####-1) as well as rat kidney<br />
and rat stomach (Order No. FA 1802-####-3).<br />
• EUROPattern from <strong>EUROIMMUN</strong> is an innovative and high-capacity automation<br />
solution for the evaluation of ANA in immunofluorescence (page 8, page 147.).<br />
EUROASSAY: Anti-ENA ProfilePlus<br />
• Differentiation of anti-nuclear antibodies (ANA).<br />
• Indications: rheumatic diseases.<br />
• Serum dilution 1 : 101; conjugate class anti-human IgG, AP-labelled.<br />
• 6 relevant anti-nuclear antibodies against “extractable nuclear antigens“ (ENA)<br />
can be detected simultaneously and monospecifically: antibodies against<br />
nRNP/Sm, Sm, SS-A, SS-B, Scl-7.0, Jo-1.<br />
• Native antigens, purified by affinity chromatography.<br />
• On request EUROASSAY can be produced with special antigen combinations,<br />
for example, with dsDNA, histones, nucleosomes, PM-Scl, nRNP/Sm, Sm, SS-A,<br />
Ro-52, SS-B, Scl-7.0, Jo-1, riboso mal P proteins, centromere protein B, M2, M4,<br />
M9, SLA/LP, LC-1, LKM-1.
<strong>EUROIMMUN</strong> Medizinische<br />
Labordiagnostika<br />
AG<br />
EUROLINE: ANA Profile 3<br />
• Differentiation of antibodies against cell nuclei (ANA).<br />
• Indications: Sharp syndrome (MCTD), systemic lupus erythematosus, Sjögren’s<br />
syndrome, systemic sclerosis, poly/dermatomyositis, PBC.<br />
• Serum dilution 1 : 101; conjugate class anti-human IgG, AP-labelled.<br />
• With the EUROLINE ANA-Profile 3, fifteen autoantibodies can be determined:<br />
anti bodies against nRNP/Sm, Sm, SS-A, Ro-52, SS-B, Scl-7.0, PM-Scl, Jo-1, centromere<br />
protein B, PCNA, dsDNA, nucleosomes, histones, ribosomal P-proteins,<br />
AMA M2.<br />
• Antibodies against SS-A are characteristic markers for SLE and Sjögren’s<br />
syndrome. In contrast, antibodies against Ro-52 also occur in patients with other<br />
autoimmune diseases.<br />
• Native antigens, purified by affinity chromatography (exception: centromere<br />
protein B, PM-Scl, Ro-52, PCNA).<br />
• Further antigen combinations: page 80.<br />
• Test strips can be automatically incubated and evaluated using the systems<br />
EUROBlotMaster und EUROLineScan (see page 27.).<br />
EUROLINE: Myositis Profile 3<br />
EUROLINE: Systemic Sclerosis Profile (Nucleoli)<br />
Autoantibodies against Cell Nuclei (ANA)<br />
• Differentiation of myositis-associated antibodies against cell-nuclear and<br />
•<br />
cytoplasmic antigens.<br />
Indications: poly/dermatomyositis.<br />
• Serum dilution 1 : 101, conjugate class anti-human IgG, AP-labelled.<br />
• With the EUROLINE Myositis Profile 3, eleven autoantibodies can be determined:<br />
anti bodies against Mi-2, Ku, PM-Scl100, PM-Scl7.5, Jo-1, SRP, PL-7., PL-12, EJ, OJ<br />
and Ro-52.<br />
• Test strips can be automatically incubated and evaluated using the systems<br />
EUROBlotMaster und EUROLineScan (see page 27.).<br />
• Differentiation of systemic sclerosis-associated antibodies against cell-nuclear<br />
antigens.<br />
• Indications: systemic sclerosis (SSc, diffuse and limited form), overlap syndromes.<br />
• Serum dilution 1 : 101, conjugate class anti-human IgG, AP-labelled.<br />
• With the EUROLINE Systemic Sclerosis Profile (Nucleoli), thirteen autoantibodies<br />
can be determined: anti bodies against Scl-7.0, CENP A, CENP B, RP11 (RNAP-III),<br />
RP155 (RNAP-III), fibrillarin, NOR-90, Th/To, PM-Scl100, PM-Scl7.5, Ku, PDGFR,<br />
Ro-52.<br />
• Test strips can be automatically incubated and evaluated using the systems<br />
EUROBlotMaster und EUROLineScan (see page 27.).<br />
nRNP/Sm<br />
Sm<br />
SS-A<br />
Ro-52<br />
SS-B<br />
Scl-7.0<br />
PM-Scl<br />
Jo-1<br />
CENP B<br />
PCNA<br />
dsDNA<br />
nucleos.<br />
histones<br />
rib. P-prot.<br />
AMA M2<br />
control<br />
Incubated EUROLINE ANA Profile 3.<br />
Order No. Formats<br />
DL 1590-1601-3 G page 82<br />
Mi-2<br />
Ku<br />
PM-Scl100<br />
PM-Scl7.5<br />
Jo-1<br />
SRP<br />
PL-7.<br />
PL-12<br />
EJ<br />
OJ<br />
Ro-52<br />
control<br />
Incubated EUROLINE Myositis Profile 3.<br />
Order No. Formats<br />
DL 1530-1601-3 G page 82<br />
Scl-7.0<br />
CENP A<br />
CENP B<br />
RP11 (RNAP-III)<br />
RP155 (RNAP-III)<br />
fibrillarin<br />
NOR-90<br />
Th/To<br />
PM-Scl100<br />
PM-Scl7.5<br />
Ku<br />
PDGFR<br />
Ro-52<br />
control<br />
Incubated EUROLINE Systemic Sclerosis Profile<br />
(Nucleoli).<br />
Order No. Formats<br />
DL 1532-1601 G page 82<br />
— 35 —<br />
<strong>EUROIMMUN</strong> <strong>Product</strong>s for the<br />
Determination of Autoantibodies
<strong>EUROIMMUN</strong> Medizinische<br />
Labordiagnostika<br />
AG<br />
<strong>EUROIMMUN</strong> <strong>Product</strong>s for the<br />
Determination of Autoantibodies<br />
Incubated ELISA ANA Screen (antigen mixture of<br />
dsDNA, histones, nRNP/Sm, Sm, SS-A, SS-B, Scl-<br />
7.0, Jo-1, riboso mal P-proteins, centro mere).<br />
Order No. Formats<br />
EA 1590-9601-8 G page 90<br />
Incubated ELISA Anti-ENA ProfilePlus 2 (antigens:<br />
riboso mal P-proteins, nRNP/Sm, Sm, SS-A, SS-B,<br />
Scl-7.0, Jo-1, centro mere).<br />
Order No. Formats<br />
EA 1590-1208-2 G page 90<br />
Incubated ELISA Anti-SS-A, Anti-SS-B.<br />
Order No. Formats<br />
EA ####-9601 G page 90<br />
— 36 —<br />
Autoantibodies against Cell Nuclei (ANA)<br />
Microplate ELISA: ANA Screen, Anti-ENA PoolPlus<br />
• Screening test for predifferentiation of antibodies against cell nuclei (ANA) and<br />
cytoplasm components.<br />
• Indications: Sharp syndrome (MCTD), systemic lupus erythematosus, Sjögren’s<br />
syndrome, progressive systemic sclerosis, polymyositis/dermatomyositis.<br />
• Serum dilution 1 : 200, conjugate class anti-human IgG, POD-labelled.<br />
• One microplate well incubated per patient.<br />
• 1-point calibration, semiquantitative.<br />
• Native antigens (exception: centromere, recombinant).<br />
• The ANA Screen ELISA supplements the gold standard immunofluorescence.<br />
It is based on a mixture of 10 highly purified antigens, which provide higher<br />
sensitivity and specificity than the undefined cell extracts used by other<br />
manufacturers.<br />
• Two ELISAs with different antigen combinations, adapted to particular indications<br />
or for follow-up of immunofluorescence patterns, are available.<br />
Microplate ELISA: SLE Profile 1/2, Anti-ENA ProfilePlus 1/2<br />
• Differentiation of antibodies against cell nuclei (ANA) and cytoplasm components.<br />
• Indications: Sharp syndrome (MCTD), systemic lupus erythematosus, Sjögren’s<br />
syndrome, progressive systemic sclerosis, polymyositis/dermatomyositis.<br />
• Serum dilution 1 : 200; conjugate class anti-human IgG, POD-labelled.<br />
• 8 or 6 relevant antibodies can be detected simultaneously.<br />
• 1-point calibration, semiquantitive. Calibrator pool and negative controls each<br />
on a separate microplate strip (SLE Profiles and Anti-ENA ProfilePlus 2) or on<br />
the same microplate strip as the patient serum.<br />
• Native antigens (exception: Ro-52, centromere and PM-Scl, recombinant).<br />
• In total 4 different ELISAs with different antigen combinations, adapted to<br />
particular indications or for follow-up of immunofluorescence patterns, are<br />
available.<br />
Microplate ELISA: ANA Single-Antigen ELISAs<br />
• Differentiation of antibodies against cell nuclei (ANA) and cytoplasm components.<br />
• Indications: rheumatic diseases.<br />
• Serum dilution 1 : 200; conjugate class anti-human IgG, POD-labelled.<br />
• Antibodies against cell nuclei components can be determined quantitatively in<br />
RU/ml.<br />
• 3-point calibration, quantitative.<br />
• Identical incubation conditions and times: all single-antigen tests can be combined<br />
with each other on one microplate.<br />
• Native antigens (exceptions: centromere and PM-Scl, recombinant).<br />
• Single-antigen ELISAs available for detection of antibodies against cell nuclei<br />
and cytoplasm antigens: ssDNA, nucleosomes, dsDNA, histones, ribosomal P<br />
proteins, PM-Scl, nRNP/Sm, Sm, SS-A, SS-B, Scl-7.0, Jo-1, centromere.
<strong>EUROIMMUN</strong> Medizinische<br />
Labordiagnostika<br />
AG<br />
Indirect Immunofluorescence Test: Crithidia luciliae<br />
• Detection of antibodies against dsDNA.<br />
• Indication: lupus erythematosus disseminatus.<br />
• Initial dilution: 1:10, conjugate class anti-human IgG, FITC-labelled.<br />
• Animal pathogenic haemoflagellates of Crithidia luciliae are used for the detection<br />
of autoantibodies against double-stranded, native DNA (dsDNA, nDNA)<br />
with indirect immunofluorescence. These protozoa possess a giant mitochondrion<br />
containing dsDNA (kinetoplast) that, in general, does not show any of the<br />
other antigens present in the cell nuclei. Antibodies reacting with the kinetoplast<br />
are therefore only directed against dsDNA.<br />
• Alongside the conventional CLIFT, which shows a particularly high disease<br />
specificity, <strong>EUROIMMUN</strong> has developed an Anti-Crithidia luciliae sensitive IFT<br />
(order no. FA 157.2-####-1), which is comparable in sensitivity to the Anti-dsDNA-<br />
NcX ELISA and the Farr assay. However, despite comparable sensitivities, the<br />
assays identify different patients. To increase the serological hit rate different<br />
test systems are often combined.<br />
Microplate ELISA: Anti-dsDNA-NcX<br />
• Monospecific detection of antibodies against dsDNA.<br />
• Indication: lupus erythematosus disseminatus.<br />
• Serum dilution 1: 200, conjugate class anti-human IgG, POD-labelled.<br />
• Antibodies against dsDNA can be determined quantitatively in IU/ml.<br />
• 3-point calibration, quantitative.<br />
• Antigen: double-stranded DNA, complexed with nucleosomes (NcX).<br />
• Due to good sensitivity and specificity, the Anti-dsDNA-NcX ELISA stands out<br />
by high diagnostic efficiency. High concentrations of autoantibodies against<br />
dsDNA in the ELISA are considered to be a reliable marker for the diagnosis<br />
or prognosis of SLE. Individual changes in the dsDNA antibody concentration<br />
correlate with the activity of the disease and can be used for monitoring the<br />
development of the disease in SLE patients. In cases of immunosuppressive<br />
therapy or clinical remission dsDNA antibodies cannot be detected with the<br />
ELISA anymore.<br />
Anti-dsDNA RIA by Farr<br />
Autoantibodies against DoubleStranded DNA (dsDNA)<br />
• Monospecific detection of antibodies against dsDNA.<br />
• ndication: lupus erythematosus disseminatus.<br />
• Use of undiluted samples.<br />
• Antigen: 125I-labelled dsDNA from plasmid DNA.<br />
• The Farr radioimmunoassay has always been of great importance. On the<br />
whole, it has the same specificity as the immunofluorescence test and the same<br />
sensitivity as the ELISA. Apparently, its well-known high diagnostic efficiency<br />
is based on the fact that only the fraction of the anti-dsDNA antibodies which<br />
is able to form bigger precipitating immune complexes with circulating DNA in<br />
liquid phase contributes to the measuring signal. The principle of the Farr test<br />
reflects, so to speak, the significant step in the pathomechanism of SLE: the<br />
formation of appropriate immune complexes, deposits of which build up in the<br />
joints, kidneys, liver and other organs.<br />
Crithidia luciliae: antibodies against dsDNA.<br />
Order No. Formats<br />
FA 157.2-#### page 132<br />
Incubated ELISA Anti-dsDNA-NcX.<br />
Order No. Formats<br />
EA 157.2-9601 G page 90<br />
RIA Anti-dsDNA.<br />
Order No. Formats<br />
RA 157.1-10001 page 93<br />
— 37. —<br />
<strong>EUROIMMUN</strong> <strong>Product</strong>s for the<br />
Determination of Autoantibodies
<strong>EUROIMMUN</strong> Medizinische<br />
Labordiagnostika<br />
AG<br />
<strong>EUROIMMUN</strong> <strong>Product</strong>s for the<br />
Determination of Autoantibodies<br />
Incubated ELISA Anti-CCP, Anti-Sa.<br />
Order No. (Anti-CCP) Formats<br />
EA 1505-9601 G page 90<br />
— 38 —<br />
<strong>EUROIMMUN</strong><br />
M edizinische<br />
Labordiagnostika<br />
AG<br />
Antibodies against cyclic citrullinated peptides (CCP):<br />
An ELISA for specifi c diagnosis of rheumatoid arthritis<br />
H<br />
N<br />
O<br />
NH<br />
H 2N + NH 2<br />
Autoantibodies against CCP and Sa<br />
Peptidylargininedeiminase<br />
(PAD)<br />
Ca 2+<br />
H<br />
N<br />
NH<br />
O NH 2<br />
L-arginine L-citrulline<br />
peroxidase-labelled<br />
anti-human antibody<br />
colourless<br />
chromogen<br />
The amino acid citrulline Principle of the anti-CCP ELISA<br />
Rheumatoid arthritis (RA) is one of the<br />
most common autoimmune diseases, affecting<br />
around 1% of the world population. It is<br />
characterised by infl ammation of the synovial<br />
membrane, which spreads symmetrically<br />
from the small to the large joints. Initial<br />
symptoms include painful swelling of fi nger<br />
joints with morning stiffness in the joints.<br />
Early diag nosis and immediate commencement<br />
of suitable therapy is necessary to<br />
keep the disease under control.<br />
The most commonly performed serological<br />
test in suspected RA cases was until now the<br />
determination of rheumatoid factors (RF).<br />
These are antibodies (predominantly of class<br />
IgM) which react with gamma globulins and<br />
occur in 60-80% of RA patients. RF are a sensitive<br />
but not very specifi c marker for RA,<br />
since they also occur in healthy individuals<br />
and in patients with various infections or<br />
other autoimmune diseases (systemic lupus<br />
erythematosus, Sjögren’s syn drome, scleroderma<br />
and others).<br />
40-60% of RA patients also exhibit autoanti<br />
bodies against epidermal fi l ag grin 1 (RA<br />
keratin, anti-pe ri nu clear fac tor) in their serum.<br />
Fil ag grin is a protein of the epi dermis,<br />
which links keratin fi laments to one another.<br />
Autoanti bodies against fi laggrin are detected<br />
by indirect immuno fl uor escence: the antigen<br />
substrate rat oesophagus shows staining of<br />
the stratum corneum (RA keratin) on the<br />
luminal side; anti-perinuclear factors (APF)<br />
are apparent in the cyto plasmic inclusion<br />
bodies of human epithelial cells of the oral<br />
mucosa.<br />
O<br />
Microplate ELISA: Anti-CCP, Anti-Sa<br />
• Screening test for the specific determination of autoantibodies against cyclic<br />
citrullinated peptides (CCP) and citrullinated Sa.<br />
• Indication: rheumatoid arthritis (RA).<br />
• Serum dilution 1 : 100; conjugate class anti-human IgG, POD-labelled.<br />
• Antibodies against CCP and Sa can be determined quantitatively in RU/ml.<br />
Optional reference control for the determination of ratio values.<br />
• Antigen: synthetic cyclic citrullinated peptides (CCP, second generation),<br />
citrullinated Sa.<br />
Antigen Order No.<br />
CCP EA 1505-9601 G<br />
Sa EA 151a-4802 G<br />
<strong>EUROIMMUN</strong> AG · 23560 Luebeck (Germany) · Seekamp 31 · Telephone +49 451 58550 · Fax 5855591 · E-mail euroimmun@euroimmun.de · www.euroimmun.com<br />
POD<br />
POD<br />
stain<br />
human antibody<br />
against CCP<br />
synthetic CCP<br />
In recent years it has been shown that the<br />
rare amino acid citrulline, which is present in<br />
fi laggrin, is a substantial component of the<br />
antigenic epitope. Enzyme immunoassays<br />
which use synthetic citrullinated peptide<br />
as the target antigen offer a useful alternative<br />
to indirect immunofl uorescence 2 . A<br />
direct comparison study demonstrated that<br />
the sensitivity can be increased from 49%<br />
to 68% by using cyclic citrullinated peptide<br />
instead of linear citrullinated peptide as an<br />
ELISA substrate 3 . Antibodies against cyc li c<br />
citrulli nated pep tides (CCP) are a new and<br />
highly specifi c marker for RA.<br />
An ti bodies against CCP are predominantly<br />
of class IgG and have a specifi city of 98%<br />
for RA. They are observed very early in the<br />
disease course and have a high predictive<br />
value: patients with anti-CCP antibodies<br />
develop signifi cantly more radiologically<br />
detectable joint damage than anti-CCPnegative<br />
patients 4 . Antibodies against CCP<br />
possess a much higher specifi city than RF<br />
(anti-CCP: 97%, RF: 62%) with the same sensitivity<br />
(anti-CCP: 79%, RF: 78%) 5 . They can<br />
be detected in early stages of the disease in<br />
79% of patients.<br />
EU ROIMMUN offers an innovative microplate<br />
ELISA for quantitative deter min ation of<br />
autoantibodies against CCP. Diluted patient<br />
sera are incubated in wells coated with synthetic<br />
cyclic citrullinated peptides (second<br />
gener ation). Specifi c antibodies in the serum<br />
bind to the immobi lised antigen and cause a<br />
photo metric colour reaction by means of an<br />
enzyme-coupled secondary antibody. Five<br />
Anti-CCP ELISA<br />
calibration sera ensure reliable measurement<br />
of antibody concentrations. The EURO-<br />
IMMUN Anti-CCP ELISA is a highly specifi c<br />
and sensitive sero logical test system for the<br />
diagnosis of RA.<br />
Panel n<br />
Anti-CCP<br />
positive<br />
Sensitivity RA 419 329 (78.5%)<br />
Asymptomatic blood<br />
donors<br />
400 2 (0.5%)<br />
Psoriatic arthritis 28 0<br />
Other arthritides 35 3 (8.6%)<br />
Systemic lupus<br />
erythematosus<br />
108 3 (2.8%)<br />
Sjögren‘s syndrome 106 2 (1.9%)<br />
Scleroderma 98 3 (3.1%)<br />
Autoimmune<br />
thyroiditis<br />
Wegener‘<br />
granulomatosis<br />
Anti-parvovirus B19<br />
positive<br />
159 4 (2.5%)<br />
25 1 (4.0%)<br />
126 3 (2.4%)<br />
Viral hepatides 54 0<br />
Anti-HIV positive 5 0<br />
Tuberculosis 10 0<br />
Specifi city RA 1154 21 (98.2%)<br />
1) Nogueira et al., Ann. Rheum. Dis. 60: 882 (2001)<br />
2) Schellekens et al., J. Clin. Invest. 101: 273-281 (1998)<br />
3) Schellekens et al., Arthritis Rheum. 43: 155-163 (2000)<br />
4) Kroot et al., Arthritis Rheum. 43: 1831-1835 (2000)<br />
5) Vasishta, Am. Clin. Lab. 21: 34-36 (2002)
<strong>EUROIMMUN</strong> Medizinische<br />
Labordiagnostika<br />
AG<br />
Autoantibodies against Mitochondria (AMA)<br />
Indirect Immunofluorescence Test: EUROPL<strong>US</strong> Rat Kidney and<br />
M2 BIOCHIPs<br />
• Screening test for the detection of antibodies against mitochondria (AMA) including<br />
simultaneous confirmation of the subtype AMA M2.<br />
• Indication: primary biliary cirrhosis (PBC).<br />
• Initial dilution 1 : 100; polyvalent conjugate anti-human IgAGM, FITC-labelled.<br />
• Rat kidney is the standard substrate for detecting anti-mitochondrial antibodies.<br />
Nine AMA types (M1 to M9) can be differentiated.<br />
• The BIOCHIP coated with M2 permits monospecific confirmation of antibodies<br />
against the native pyruvate dehydrogenase complex and the recombinant M2<br />
fusion protein (BPO) in one single test procedure, thus a PBC can be diagnosed<br />
serologically with con fidence.<br />
• This BIOCHIP Mosaic can be supplemented as required using additional<br />
substrates, e.g. HEp-2 cells (ANA, nuclear dots), rat liver (liver-kidney microsomes,<br />
LKM) or rat stomach (ASMA).<br />
EUROASSAY: AMA Profile M2, M4, M9<br />
• Differentiation of mitochondrial antibodies (AMA).<br />
• Indication: primary biliary cirrhosis (PBC).<br />
• Serum dilution 1 : 100; conjugate class anti-human IgGM, AP-labelled.<br />
• 3 relevant mitochondrial antibodies can be detected simultaneously and monospecifically:<br />
antibodies against M2, M4, M9.<br />
• Native antigens: pyruvate dehydrogenase complex (M2), sulfite oxidase (M4),<br />
gly cogen phosphorylase (M9).<br />
Anti- Associated diseases<br />
Rat kidney and M2 BIOCHIP: antibodies against<br />
mitochondria (AMA).<br />
M2 Primary biliary cirrhosis (high titers), other chronic liver diseases<br />
M4 Primary biliary cirrhosis<br />
M9 Early phase of primary biliary cirrhosis Incubated EUROASSAY AMA Profile M2, M4, M9.<br />
Microplate ELISA: Anti-M2-3E<br />
• Differentiation of mitochondrial antibodies (AMA).<br />
• Indication: primary biliary cirrhosis (PBC).<br />
• Serum dilution 1 : 100; conjugate class anti-human IgG, POD-labelled.<br />
• Antibodies against M2 can be determined quantitatively in RU/ml.<br />
• Antigen: native pyruvate dehydrogenase complex plus recombinant M2 fusion<br />
protein (BPO) containing the immunogenic domains of the E2 subunits of PDH,<br />
BCOADH and OGDH.<br />
Order No. Formats<br />
FA 1620-####-3 page 132<br />
Order No. Formats<br />
DA 1620-####-1 O page 80<br />
Incubated ELISA anti-M2-3E.<br />
Order No. Formats<br />
EA 1622-9601 G page 91<br />
— 39 —<br />
<strong>EUROIMMUN</strong> <strong>Product</strong>s for the<br />
Determination of Autoantibodies
<strong>EUROIMMUN</strong> Medizinische<br />
Labordiagnostika<br />
AG<br />
<strong>EUROIMMUN</strong> <strong>Product</strong>s for the<br />
Determination of Autoantibodies<br />
HEp-2 cells: anti-nuclear dots. Primate liver:<br />
anti-LSP. Rat kidney: AMA. Rat liver: ANA. Rat<br />
stomach: ASMA. VSM47.: Anti-F-actin.<br />
Order No. Formats<br />
FA 1300-####-8 page 126<br />
Rat liver and rat kidney: antibodies against liverkidney<br />
microsomes (anti-LKM).<br />
Order No. Formats<br />
FA 1300-####-1 page 126<br />
— 40 —<br />
Autoantibodies against Liver Antigens<br />
Indirect Immunofluorescence Test: Liver Mosaic 8<br />
• Screening and differentiation test for the detection of liver-specific antibodies,<br />
antibodies against mitochondria (AMA), antibodies against cell nuclei (ANA),<br />
antibodies against smooth muscles (ASMA), F-actin and other autoantibodies.<br />
• Indications: autoimmune hepatitis, primary biliary cirrhosis, rheumatic diseases.<br />
• Initial dilution 1 : 100; polyvalent conjugate anti-human IgAGM, FITC-labelled.<br />
• The BIOCHIP Mosaic consists of 6 substrates: human epithelial cells (HEp-2),<br />
primate liver, rat kidney, rat liver, rat stomach, VSM47.. Thus, a broad spectrum<br />
of antigens is present, allowing not only targeted serological diagnoses, but also<br />
frequently yielding additional results with cli nical relevance.<br />
• Antibodies against cell nuclei (ANA) can be particularly well demonstrated using<br />
HEp-2 cells and primate liver, and are identified according to their fluorescence<br />
patterns. However, they also stain the cell nuclei of the other tissues more or<br />
less intensely. Clinical significance: rheumatic diseases, primary biliary cirr hosis<br />
(antibodies against nuclear dots).<br />
• With primate liver, several liver-specific autoantibodies can be investigated e.g.<br />
anti bodies against liver cell membrane (anti-LMA) and liver-specific protein<br />
(anti-LSP). Clinical significance: autoimmune hepa titis.<br />
• Antibodies against mitochondria (AMA) show a granular cytoplasmic fluorescence<br />
on all 6 substrates. With the standard substrate rat kidney, the proximal<br />
and distal tubule cells fluoresce equally. Clinical significance: primary biliary<br />
cirrhosis.<br />
• Autoantibodies against liver-kidney microsomes (anti-LKM) react with rat liver<br />
and rat kidney (see below). The other substrates are essentially negative.<br />
• In the case of autoantibodies against smooth muscles (ASMA), the tunica<br />
muscularis, the lamina muscularis mucosa as well as the interglandular contractile<br />
fibrils fluoresce on the rat stomach. ASMA directed against the target<br />
antigen F-actin furthermore react with the cytoskeleton of HEp-2 cells and the<br />
bile canaliculi of primate liver. The substrate VSM47. reacts very specifically,<br />
showing a filamentous, needle-like fluorescence. Clinical signifi cance:<br />
•<br />
autoimmune (lupoid) chronic active hepatitis.<br />
The BIOCHIP Mosaic can be supplemented as required with additional substrates,<br />
e.g. Crithidia luciliae (antibodies against dsDNA), musculus iliopsoas<br />
(antibodies against skeletal muscles), heart (anti bodies against striated muscles,<br />
antibodies against intercalated disks, AMA M7.), different EUROPL<strong>US</strong> substrates<br />
(AMA-M2-3E, Sp100, gp210, PML, SLA/LP, LC-1, LKM).<br />
Indirect Immunofluorescence Test: BIOCHIP Mosaic Rat Liver/<br />
Rat Kidney (Liver Mosaic 1)<br />
• Specific detection of antibodies against liver-kidney microsomes (anti-LKM).<br />
• Indications: autoimmune hepatitis, often associated with extrahepatic syndromes<br />
such as arthralgias, glomerulonephritis, vitiligo and chronic inflammatory bowel<br />
diseases.<br />
• Initial dilution 1 : 100; polyvalent conjugate anti-human IgAGM, FITC-labelled.<br />
• Autoantibodies against liver-kidney microsomes react with rat liver and generate<br />
a smooth staining in the cytoplasm of the hepatocytes.<br />
• In rat kidney, particularly in the cortex area, a fine granular fluorescence of<br />
the proximal tubules – recognizable by the luminal brush border – is visible.<br />
The distal tubules are negative. The fluorescence intensity of the liver cells is<br />
normally stronger than that of the proximal renal tubules.<br />
• The differentiation between autoimmune hepatitis and virus-induced hepatitis<br />
can additionally be accomplished by investigating the appropriate viral<br />
parameters.
<strong>EUROIMMUN</strong> Medizinische<br />
Labordiagnostika<br />
AG<br />
EUROLINE: Profile Autoimmune Liver Diseases<br />
• Differentiation of antibodies in autoimmune liver diseases.<br />
• Indications: autoimmune hepatitis, primary biliary cirrhosis, overlap syndromes.<br />
• Serum dilution 1 : 100, conjugate class anti-human IgG, AP-labelled.<br />
• With the EUROLINE Profile Autoimmune Liver Diseases, nine autoantibodies can<br />
be determined: anti bodies against AMA M2, M2-3E (BPO), Sp100, PML, gp210,<br />
LKM-1, LC-1, SLA/LP and Ro-52.<br />
• Test strips can be automatically incubated and evaluated using the systems<br />
EUROBlotMaster und EUROLineScan (see page 27.).<br />
• Further antigen combinations on page 82.<br />
Anti- Associated Diseases<br />
M2, Sp100, PML, gp210 Primary biliary cirrhosis<br />
LKM-1, SLA/LP, LC-1 Autoimmune hepatitis<br />
Ro-52 Autoimmune hepatitis, rheumatic diseases<br />
EUROASSAY: Liver Profile<br />
(Anti-M2, -LKM-1, LC-1, -SLA/LP)<br />
Autoantibodies against Liver Antigens<br />
• Determination of mitochondrial antibodies AMA M2, antibodies against liverkidney<br />
microsomes type 1 (LKM-1), antibodies against liver cytosolic antigen<br />
type 1 (LC-1), as well as of antibodies against soluble liver antigen/liver-pancreas<br />
antigen (SLA/LP).<br />
• Indication: autoimmune liver diseases.<br />
• Serum dilution 1 : 100, conjugate class anti-human IgG, AP-labelled.<br />
• Antibodies against M2, LKM-1, LC-1 and SLA/LP can be detected simultaneously<br />
and mono specifically.<br />
• Antigens: pyruvate dehydrogenase complex (M2, native), cytochrome P450 IID6<br />
(LKM-1, recombinant), formiminotransferase-cyclodeaminase (LC-1, recombinant)<br />
and soluble liver antigen/liver-pancreas antigen (SLA/LP, recombinant).<br />
Microplate ELISA: Anti-SLA/LP, Anti-LC-1, Anti-LKM-1<br />
3E (BPO)<br />
Sp100<br />
PML<br />
gp210<br />
LKM-1<br />
LC-1<br />
SLA/LP<br />
Ro-52<br />
Incubated EUROLINE Profile Autoimmune Liver<br />
Diseases.<br />
Incubated EUROASSAY M2, LKM-1, LC-1, SLA/LP.<br />
• Monospecific determination of antibodies against soluble liver antigen/liverpancreas<br />
antigen (SLA/LP), cytosolic liver antigen type 1 (LC-1) and liver-kidney<br />
microsomes type 1 (LKM-1).<br />
• Indication: autoimmune hepatitis.<br />
• Serum dilution 1 : 100, conjugate class anti-human IgG, POD-labelled.<br />
• 3-point calibration, quantitative (exception: Anti-LC-1, semi-quantitative).<br />
• Identical incubation conditions and times: all tests can be combined without<br />
difficulty on one and the same microplate.<br />
• Recombinant antigens: soluble liver antigen/liver-pancreas antigen (SLA/LP),<br />
formiminotransferase-cyclodeaminase (LC-1) and cytochrome P450 IID6 (LKM-<br />
1). The corresponding human cDNA was expressed in E. coli (SLA/LP) or insect<br />
cells (LC-1, LKM-1). Incubated ELISA Anti-SLA/LP, Anti-LC-1, Anti-LKM-1.<br />
AMA M2<br />
control<br />
Order No. Formats<br />
DL 1300-1601-4 G page 82<br />
Order No. Formats<br />
DA 1300-1003-3 G page 80<br />
Order No. (Anti-SLA/LP) Formats<br />
EA 1302-9601 G page 89<br />
— 41 —<br />
<strong>EUROIMMUN</strong> <strong>Product</strong>s for the<br />
Determination of Autoantibodies
<strong>EUROIMMUN</strong> Medizinische<br />
Labordiagnostika<br />
AG<br />
<strong>EUROIMMUN</strong> <strong>Product</strong>s for the<br />
Determination of Autoantibodies<br />
— 42 —<br />
Autoantibodies against Thyroid gland Antigens / Antigen Detections<br />
Thyroid gland, unfixed and thyroglobulin<br />
BIOCHIP: anti bodies against thyroid microsomes<br />
and thyro globulin.<br />
Order No. Formats<br />
FA 1010-####-3 page 121<br />
RIA for thyroid diagnostics.<br />
Order No. (Anti-TPO) Formats<br />
RA 1012-####-# page 93<br />
Incubated ELISA Anti-Thyroid antigens.<br />
Order No. (Anti-TPO) Formats<br />
EA 1012-9601 G page 89<br />
Indirect Immunofluorescence Test: EUROPL<strong>US</strong> Thyroid Gland<br />
(unfixed) and Thyroglobulin<br />
• Detection of antibodies against thyroid gland antigens.<br />
• Indications: Basedow’s disease, Hashimoto autoimmune thyroiditis.<br />
• Initial dilution 1 : 10; conjugate class anti-human IgG, FITC-labelled.<br />
• Using unfixed thyroid tissue, two important thyroid-specific antibodies can<br />
be found: Autoantibodies against thyroid microsomes (MAb) give a granular<br />
staining in the cytoplasm of the follicle epithelium (target antigen: thyroid<br />
peroxidase, TPO). Autoantibodies against thyroglobulin (TAb) react with the<br />
colloid of all follicles of the thyroid tissue and cause a reticular fluorescence<br />
pattern.<br />
• With the thyroglobulin-coated BIOCHIP, autoantibodies against thyroglobulin<br />
(TG) can be confirmed monospecifically in one and the same test procedure.<br />
• This BIOCHIP Mosaic can be supplemented as required with further substrates,<br />
e.g. rat kidney, to achieve a differentiation of antibodies against thyroid microsomes<br />
from mitochondrial antibodies (AMA). For a serological diagnosis of autoimmune<br />
polyendocrinopathies, BIOCHIPs with frozen sections of pancreas,<br />
adrenal gland, ovary, testis and stomach can be added.<br />
Radioimmunoassays (RIA/IRMA): Thyroid Specific Autoantibodies,<br />
Antigens and Hormones<br />
• Monospecific detection of autoantibodies against thyroglobulin (TG), thyroid<br />
peroxidase (TPO) and thyrotropin receptor (TSH-R).<br />
• Specific detection of the thyroid antigen thyrotropin and the hormones free<br />
triiodothyronine (FT3), free thyroxine (FT4), thyrotropin (TSH), calcitonin.<br />
• Indications: Basedow’s disease, Hashimoto autoimmune thyroiditis, medullary<br />
thyroid carcinoma, thyroidal C-cell hyperplasia, therapy controls in hyper- and<br />
hypothyrosis.<br />
• Serum dilutions: 1:50 for anti-TG and anti-TPO (magnetic), 1:20 for anti-TG and<br />
anti-TPO (precipitation), undiluted for all remaining test kits.<br />
• 5-point to 8-point calibration (quantitative).<br />
Analyte Order No. Analyte Order No.<br />
Anti-TPO RA 1012-####-# FT3 RD 1016-10001<br />
Anti-TG RA 1013-10001-# FT4 RD 1017.-10001<br />
TRAb RA 1015-10001-# TSH RD 1018-10001<br />
Thyroglobulin RD 1013-10001 Calcitonin RD 1019-10001<br />
Microplate ELISA: Anti-Thyroglobulin, Anti-Thyroid Peroxidase,<br />
Anti-TSH receptor<br />
• Monospecific determination of antibodies against thyroglobulin (TG), thyroid<br />
peroxidase (TPO), and thyrotropin receptor (TSH-R).<br />
• Indications: Basedow’s disease, Hashimoto autoimmune thyroiditis.<br />
• Serum dilution 1 : 200 (exception: anti-TSH-R undiluted); conjugate class antihuman<br />
IgG, POD-labelled (anti-TSH-R: avidin-labelled).<br />
• 3-point calibration (exception: anti-TSH-R, 5-point calibration).<br />
• The quantification is carried out according to international reference preparations<br />
(anti-TG: NIBSC 65/93; anti-TPO: NIBSC 66/387.; anti-TSH-R: NIBSC 90/67.2).<br />
• Thyroglobulin/TSH-R: native antigen; thyroid peroxidase: recombinant antigen.<br />
Antigen Order No.<br />
Thyroid peroxidase EA 1012-9601 G<br />
Thyroglobulin EA 1013-9601 G<br />
TSH receptor EA 1015-9601 G<br />
TSH receptor (Fast-ELISA) EA 1015-9601-1 G
<strong>EUROIMMUN</strong> Medizinische<br />
Labordiagnostika<br />
AG<br />
Indirect Immunofluorescence test: Dermatology Mosaic 7.<br />
• Screening and differentiation test for detection of skin-specific antibodies.<br />
• Indication: bullous autoimmune dermatoses.<br />
Autoantibodies against Antigens of the Skin<br />
• Initial dilution 1 : 10; conjugate class anti-human IgG, FITC-labelled.<br />
• The BIOCHIP Mosaic consists of 6 substrates: primate oesophagus, salt-split skin,<br />
desmoglein-1-expressing cells, desmoglein-3-expressing cells, BP230-expressing<br />
cells (gc) and BP180 (EUROPL<strong>US</strong>). Thus a comprehensive antigen spectrum is<br />
available in a single analysis, allowing targeted serological diagnosis.<br />
• Antibodies against prickle cell desmosomes react with surface antigens of<br />
keratinocytes. Tissue sections of oesophagus and tongue show a granular<br />
fluo rescence of the intercellular matter in the whole stratum spinosum, but<br />
differentiation between desmoglein 1 and desmoglein 3 is difficult. When<br />
specific transfected cells are employed in addition, a targeted diagnosis in a<br />
single test run is possible.<br />
• Antibodies against basal membrane structures react with salt-split skin. Anti-<br />
BP180, anti-BP230 and anti–LAD97 cause staining of the epidermal side, and<br />
antibodies against laminin 5, collagen VII and other antigens staining of the<br />
dermal side of salt-split skin.<br />
• When autoantibodies against BP180 or BP230 are present, the epidermal basal<br />
membrane in the oesophagus or tongue is visible as a fine linear colouring<br />
between the stratum basale and the connective tissue. These antibodies can<br />
be differentiated by means of BP180-NC16A-4X coated BIOCHIPs and cells<br />
specifically transfected with BP230 (globular C-terminal domain (gC)).<br />
• This BIOCHIP Mosaic can be customised with further substrates if required,<br />
e.g. tongue (antibodies against prickle cell desmosomes, epidermal basal<br />
membrane), bladder (antibodies against plakins).<br />
Substrate Order No.<br />
Oesophagus FA 1501-####<br />
Oesophagus / Tongue FA 1501-####-1<br />
Tongue FA 1502-####<br />
Bladder mucosa FA 1507.-####<br />
Salt split skin FA 150b-####<br />
BP180-NC16A-4X / BP230gC FA 1502-####-1<br />
Envoplakin FA 1491-####-50<br />
Collagen type VII NC1 FA 1947.-####-50<br />
Microplate ELISA: Anti-Desmoglein 1, Anti-Desmoglein 3, Anti-<br />
Envoplakin, Anti-BP180-NC16A-4X, Anti-BP230-CF<br />
• Monospecific detection of antibodies against desmoglein 1, desmoglein 3, envoplakin,<br />
BP180 und BP230.<br />
• Indication: bullous autoimmune dermatoses.<br />
• Serum dilution 1 : 100; conjugate class anti-human IgG, POD-labelled.<br />
• 3-point calibration, quantitative (exception: anti-envoplakin, semiquantitative).<br />
• Identical incubation conditions and times: all tests can be combined on one<br />
microplate.<br />
• Recombinant antigens: extracellular domain of desmoglein 1 or 3, envoplakin,<br />
tetramer of NC16A domain of BP180 protein, C-terminal segment of BP230<br />
protein. The corresponding human cDNA is produced in E. coli (BP180-NC16A-<br />
4X, BP230-CF, envoplakin) or in mammalian cells (desmoglein 1, desmoglein 3).<br />
Oesophagus and salt-split skin (top left, middle<br />
left): ab against epid. basal membrane. BP230<br />
gc (top right). BP180 (NC16A-4X) (middle right).<br />
Desmoglein 1 + 3 (bottom left and right).<br />
Order No. Formats<br />
FA 1501-####-7. page 128<br />
Incubated ELISA Anti-Desmoglein 1 and 3, Anti-<br />
BP180-NC16A-4X, Anti-BP230-CF.<br />
Order No. (Anti-Dsg-1) Formats<br />
EA 1495-4801 G page 89<br />
— 43 —<br />
<strong>EUROIMMUN</strong> <strong>Product</strong>s for the<br />
Determination of Autoantibodies
<strong>EUROIMMUN</strong> Medizinische<br />
Labordiagnostika<br />
AG<br />
<strong>EUROIMMUN</strong> <strong>Product</strong>s for the<br />
Determination of Autoantibodies<br />
Cerebellum: antibodies against GAD and Yo (top),<br />
against Hu and Ri (middle). Peripheral nerves:<br />
anti bodies against myelin and MAG (bottom).<br />
Order No. Formats<br />
FA 1111-####-1 page 122<br />
— 44 —<br />
amphiphysin<br />
CV2<br />
PNMA2 (Ma2/Ta)<br />
Ri<br />
Yo<br />
Hu<br />
recoverin<br />
SOX1<br />
titin<br />
control<br />
Incubated EUROLINE Neuronal Antigens Profile 4.<br />
Order No. Formats<br />
DL 1111-1601-4 G page 82<br />
Autoantibodies against Neuronal Antigens<br />
Indirect Immunofluorescence Test: Neurology Mosaic 1<br />
• Screening test for the detection of antibodies against neuronal antigens.<br />
• Indications: neurological diseases.<br />
• Initial dilution 1 : 10; conjugate class anti-human IgAGM, FITC-labelled.<br />
• Primate cerebellum and primate nerves are the standard substrates for the<br />
determination of various neuronal antibodies. The parallel use of primate<br />
intestine permits the reliable differentiation from other autoantibodies (e.g.<br />
ANA) and makes it possible to distinguish between anti-Ri and anti-Hu.<br />
• Antibodies against grey matter react intensively with the stratum granulosum<br />
and in a weaker form with the stratum moleculare of the cerebellum. Target<br />
antigen: glutamic acid decarboxylase (GAD). Clinical significance: stiff person<br />
syndrome, dia betes mellitus type I.<br />
• Antibodies against Yo stain exclusively the cytoplasm of the Purkinje cells in the<br />
cerebellum. Clinical significance: paraneoplastic neurological syndromes (PNS),<br />
indi cation of a malignoma.<br />
• In the case of antibodies against Hu and Ri all neurone nuclei in the grey matter<br />
show a granular fluorescence. Hu antibodies react in the intestine with cell nuclei<br />
of the plexus myentericus, whereas Ri antibodies do not. Clinical significance:<br />
paraneoplastic neurological syndromes (PNS), indication of a malignoma.<br />
• The white matter of the cerebellum is stained by antibodies against myelin,<br />
which present as hyaline cylinders in tissue sections of peripheral nerves. A<br />
“droplike“ ring-shaped fluorescence is observed in cross sections of nerves.<br />
• The fluorescence of the Virchow-Robin space (cerebellum, optic nerve) and the<br />
pia is caused by autoantibodies developed in neuromyelitis optica (NMO-IgG).<br />
• Antibodies against myelin-associated glycoprotein (MAG), on the other hand,<br />
show a streaky fluorescence pattern on nerve tissue and a mostly fine-granular<br />
ring-shaped fluorescence on cross sections of peripheral nerves. Clinical<br />
significance: paraproteinaemic neuropathy.<br />
• The BIOCHIP Mosaic can be supplemented as required with further substrates,<br />
e.g. cerebrum (antibodies against astrocytes), optic nerve, primate liver plus<br />
HEp-2 cells (to rule out ANA), Crithidia luciliae (anti-dsDNA), primate stomach<br />
(parietal cell antibodies), Borrelia (neuroborreliosis-associated). Aquaporin-<br />
4(AQP4) transfected HEK cells allow a monospecific antibody determination in<br />
suspected cases of neuromyelitis optica (NMO).<br />
EUROLINE: Neuronal Antigens Profile 4<br />
• Differentiation of antibodies against neuronal antigens.<br />
• Indication: paraneoplastic neurologic syndromes (PNS).<br />
• Serum dilution 1 : 100; conjugate class anti-human IgG, AP-labelled.<br />
• With the EUROLINE Neuronal Antigens Profile 4, nine autoantibodies can be<br />
determined: anti bodies against amphiphysin, CV2, PNMA2 (Ma2/Ta), Ri, Yo, Hu,<br />
recoverin, SOX1 and titin.<br />
• Test strips can be automatically incubated and evaluated using the systems<br />
EUROBlotMaster und EUROLineScan (see page 27.).
<strong>EUROIMMUN</strong> Medizinische<br />
Labordiagnostika<br />
AG<br />
Autoantibodies against Neuronal Antigens<br />
Indirect Immunofluorescence Test: Autoimmune Encephalitis<br />
Mosaic 1<br />
• Screening test for detection of neuropil antibodies (glutamate receptors type<br />
NMDA and type AMPA, LGI1, CASPR2, GABA receptors).<br />
B<br />
• Indication: autoimmune encephalitis.<br />
• Initial dilution 1 : 10; conjugate class anti-human IgG, FITC-labelled.<br />
• Immunohistochemistry with tissue sections of rat hippocampus and rat<br />
•<br />
cerebellum allows identification of antibodies against glutamate receptors (type<br />
NMDA, type AMPA) and other antibodies (e.g. VGKC-associated proteins LGI1<br />
and CASPR2). The parallel use of transfected HEK293 cells enables sensitive<br />
and monospecific antibody detection and differentiation of various neuropil<br />
antibodies.<br />
Neuropil antibodies show a flat, smooth to fine-granular fluorescence in the<br />
cytoplasm of transfected HEK293 cells. Clinical significance: autoimmune<br />
encephalitis.<br />
Transfected cells: antibodies against NMDAR<br />
and CASPR2 (top), AMPAR1 and LGI1 (middle),<br />
AMPAR2 and GABA B -R (bottom).<br />
Order No. Formats<br />
FA 112d-####-1 page 123<br />
— 45 —<br />
<strong>EUROIMMUN</strong> <strong>Product</strong>s for the<br />
Determination of Autoantibodies
<strong>EUROIMMUN</strong> Medizinische<br />
Labordiagnostika<br />
AG<br />
<strong>EUROIMMUN</strong> <strong>Product</strong>s for the<br />
Determination of Autoantibodies<br />
Primate pancreas: antibodies against islet cells.<br />
Order No. Formats<br />
FA 1020-#### page 121<br />
Incubated ELISA anti-GAD.<br />
Order No. (Anti-GAD) Formats<br />
EA 1022-9601 G page 89<br />
RIA Anti-IA2.<br />
Order No. (Anti-IA2) Formats<br />
RA 1023-#### page 93<br />
— 46 —<br />
Autoantibodies against Islet Cell Antigens<br />
Indirect Immunofluorescence Test: Primate Pancreas<br />
• Detection of antibodies against islet cells.<br />
• Indications: Differentiation between a late manifestation of diabetes type 1<br />
(latent autoimmune diabetes in adulthood, LADA) and diabetes type 2.<br />
• For a reliable determination of antibodies against islet cells an extended<br />
incubation time of 18 hours for the patient serum must be observed. The<br />
incubation time may be reduced to 2 hours but this will lead to a decrease in<br />
the sensitivity of the antibody detection test.<br />
• Standardised control with JDF units available (order no. CA 1021-0101-1).<br />
• With indirect immunofluorescence autoantibodies against pancreas islets<br />
•<br />
(ICA) can be detected in 80% of patients with new-onset diabetes type 1. Two<br />
target antigens of ICA have been identified so far: the enzymes glutamic acid<br />
decarboxylase (GAD) and tyrosine phosphatase (IA2).<br />
This BIOCHIP may be supplemented with further substrates, e. g. primate<br />
cerebellum for the detection of antibodies against GAD.<br />
• The microscopic evaluation can be significantly simplified by using small<br />
BIOCHIPs (1 x 1 mm). The BIOCHIPs appear almost completely in the field<br />
of view and facilitate finding the islet cells, thus rendering a time-consuming<br />
search unnecessary, especially in negative samples.<br />
Microplate ELISA: Anti-GAD, Anti-IA2, Anti-GAD/IA2 Pool<br />
• Monospecific detection of antibodies against glutamic acid decarboxylase (GAD),<br />
tyrosine phosphatase (IA2) or bispecific detection of both antibodies in a single<br />
reagent well.<br />
• Indications: early diagnosis of diabetes mellitus type 1, risk prediction in<br />
first grade relatives, prognosis of the clinical progression of diabetes type 1<br />
for prediction of insulin dependence, differential diagnosis in gestational<br />
diabetes, differentiation between a late manifestation of diabetes type 1 (latent<br />
autoimmune diabetes in adulthood, LADA) and diabetes type 2.<br />
• Use of undiluted samples. Similar incubation conditions and times. Manual or<br />
automated test performance.<br />
• Multipoint calibration. The quantitation is based on an international reference<br />
preparation (NIBSC 97./550).<br />
• GAD and IA2: human, recombinant antigens.<br />
Antigen Order no.<br />
Glutamic acid decarboxylase (GAD) EA 1022-9601 G<br />
Tyrosine phosphatase (IA2) EA 1023-9601 G<br />
GAD/IA2 Pool EA 1022-9601-1 G<br />
RIA: Anti-GAD, Anti-IA2, Anti-Insulin<br />
• Monospecific detection of antibodies against glutamic acid decarboxylase (GAD),<br />
tyrosine phosphatase (IA2) and insulin.<br />
• Indications: Early diagnosis of diabetes mellitus type 1, risk prediction in<br />
first grade relatives, prognosis of the clinical progression of diabetes type 1<br />
for prediction of insulin dependence, differen tial diagnosis in gestational<br />
diabetes, differentiation between a late manifestation of diabetes type 1 (laten t<br />
autoimmune diabetes in adulthood, LADA) and diabetes type 2.<br />
• Use of undiluted samples. Similar incubation conditions and times. Manual or<br />
automated test performance.<br />
• Test kit formats for 50 or 100 determinations.<br />
• GAD and IA2: human, recombinant, 125 I-labelled antigens, insulin: human,<br />
synthetic, 125 I-labelled antigen.<br />
Antigen Order no.<br />
Glutamat-Decarboxylase (GAD) RA 1022-####<br />
Tyrosin-Phosphatase (IA2) RA 1023-####<br />
Insulin RA 1024-####
<strong>EUROIMMUN</strong> Medizinische<br />
Labordiagnostika<br />
AG<br />
Indirect Immunfluorescence Test: Primate Stomach with Urea<br />
Pretreatment<br />
• Screening test for detection of antibodies against parietal cells.<br />
• Indications: forms of chronic atrophic gastritis, pernicious anemia, funicular<br />
myelosis, various autoimmune endocrinopathies such as Basedow’s and<br />
Addison’s diseases.<br />
• Initial dilution 1 : 10; polyvalent conjugate anti-human IgG, FITC-labelled.<br />
• Primate stomach is the standard substrate for detection of parietal cell antibodies.<br />
For titration, stomach tissue from rat or mouse is sufficient.<br />
• With positive results the parietal cells show a course granular to clumpy<br />
fluorescence, and the surrounding areas are usually dark.<br />
• Parietal cell antibodies (PCA) are often mixed up with antibodies against<br />
mitochondria (AMA) in microscopic analysis. The latter give an even fine<br />
granular fluorescence of the parietal cell cytoplasm, with the surrounding region<br />
showing a (weaker) reaction.<br />
• For reliable differentiation of both types of antibody, a 30-minute pretreatment<br />
of the tissue sections with urea-glycine buffer (order no. ZF 1140-0101, see page<br />
149) is recommended.<br />
• The cytomplasmic fluorescence of parietal cells resulting from PCA occurs with<br />
the same intensity with or without urea pretreatment. The typical pattern of<br />
mitochondrial antibodies is almost completely supressed by urea pretreatment,<br />
greatly facilitating PCA diagnostics.<br />
• In some AMA-positive samples it is possible to detect PCA that are obscured by<br />
the AMA pattern in conventional tissue sections.<br />
• The urea-pretreated tissue shows a significantly darker background, enabling<br />
specific fluorescence to be more easily and reliable identified.<br />
• This BIOCHIP can be supplemented with additional substrates, for example,<br />
thyroid (thyroid peroxidase, thyroglobulin), pancreas (pancreas islets), adrenal<br />
gland (adrenal cortex), ovary (ovary antigens), testis (Leydig cells), and intrinsic<br />
factor.<br />
Microplate ELISA: Anti-Parietal Cells<br />
Autoantibodies against Parietal Cells (PCA)<br />
• Monospecific detection of antibodies against parietal cells (PCA).<br />
• Indications: forms of chronic atrophic gastritis, pernicious anemia, funicular<br />
myelosis, various autoimmune endocrinopathies such as Basedow’s and<br />
Addison’s diseases<br />
• Serum dilution 1 : 200; conjugate class anti-human IgG, POD-labelled.<br />
• 3-point calibration, quantitative.<br />
• Native antigen: H + /K + -ATPase, purified by affinity chromatography.<br />
Primate stomach: antibodies against parietal<br />
cells. With urea-pretreatment (top) and without<br />
urea-pretreatment (bottom).<br />
Primate stomach: antibodies against mitochondria.<br />
With urea-pretreatment (top) and without<br />
urea-pretreatment (bottom).<br />
Order No. Formats<br />
FA 1360-#### page 126<br />
Incubated ELISA Anti-Parietal Cells.<br />
Order No. Formats<br />
EA 1361-9601 G page 89<br />
— 47. —<br />
<strong>EUROIMMUN</strong> <strong>Product</strong>s for the<br />
Determination of Autoantibodies
<strong>EUROIMMUN</strong> Medizinische<br />
Labordiagnostika<br />
AG<br />
<strong>EUROIMMUN</strong> <strong>Product</strong>s for the<br />
Determination of Autoantibodies<br />
— 48 —<br />
Autoantibodies against granulocyte Cytoplasm (cANCA/pANCA)<br />
EUROPL<strong>US</strong> Granulocyte Mosaic 25 (granuloc.<br />
(EOH), granuloc. (HCHO), PR3, MPO, GBM, HEp-2<br />
+ granuloc. (EOH)): pANCA.<br />
Order No. Formats<br />
FA 1201-####-25 page 125<br />
Incubated ELISA ANCA Profile (antigens: proteinase<br />
3, lactoferrin, myeloperoxidase, elastase,<br />
cathepsin G, BPI.<br />
Order No. Formats<br />
EA 1200-1208-1 G page 89<br />
Indirect Immunofluorescence Test: EUROPL<strong>US</strong> Granulocyte<br />
Mosaic 25<br />
• Screening test for the detection of antibodies against granulocyte cytoplasm<br />
(ANCA).<br />
• Indications: Wegener‘s granulomatosis, various forms of glomerulonephritis,<br />
primary sclerosing cholangitis, ulcerative colitis, Crohn’s disease.<br />
• Initial dilution: serum 1 : 10; conjugate anti-human IgG, FITC-labelled.<br />
• Using ethanol-fixed granulocytes, antibodies against granulocyte cytoplasm can<br />
be detected. In this case, two fluorescence patterns have to be differentiated:<br />
a granular fluorescence which is distributed evenly over the entire cytoplasm,<br />
leaving the cell nuclei free (cyto plasmic type, cANCA) or a smooth fluorescence<br />
wrapped ribbon-like around the cell nuclei (perinuclear type, pANCA).<br />
• Antibodies against all relevant granulocyte antigens as well as against further,<br />
as yet unknown antigens are detected simultaneously:<br />
Pattern Target antigen Associated diseases<br />
cANCA Proteinase 3 Wegener‘s granulomatosis<br />
pANCA Myeloperoxidase Microscopic arteritis, Churg-Strauss syndrome,<br />
polyarteritis nodosa<br />
pANCA Elastase Ulcerative colitis, Crohn’s disease, primary sclerosing<br />
cholangitis, systemic lupus erythema tosus<br />
pANCA Cathepsin G Ulcerative colitis, primary sclerosing cholangitis,<br />
Crohn’s disease<br />
pANCA Lysozyme Ulcerative colitis, primary sclerosing cholangitis,<br />
Crohn’s disease<br />
pANCA Lactoferrin Ulcerative colitis, primary sclerosing cholangitis,<br />
Crohn’s disease, systemic lupus erythema tosus,<br />
rheumatoid arthritis<br />
cANCA BPI Primary sclerosing cholangitis, ulcerative colitis,<br />
or pANCA Crohn’s disease<br />
pANCA unknown Ulcerative colitis, Crohn’s disease<br />
• The composite substrate HEp-2 cells + granulocytes enables one to differentiate<br />
between pANCA and anti-nuclear antibodies (ANA) which can easily be confused<br />
when using ethanol-fixed granu lo cytes: In the case of a positive ANA result all<br />
nuclei of the HEp-2 cells fluoresce, whereas in the case of pANCA (as well as<br />
cANCA) only the granulocytes fluoresce.<br />
• The EUROPL<strong>US</strong> substrates PR3 and MPO as monospecific tests can confirm<br />
results from conventional granulocyte screening tests. Recombinant GBM<br />
EUROPL<strong>US</strong> substrate also provides additional reliability for diagnosis. When<br />
fluorescence patterns are unclear (e.g. unspecific fluorescence caused by other<br />
cytoplasmic antibodies) these substrates facilitate evaluation.<br />
Microplate ELISA: ANCA Profile<br />
• Differentiation of antibodies against granulocyte cytoplasm (ANCA).<br />
• Indications: Wegener‘s granulomatosis, various forms of glomerulonephritis,<br />
primary sclerosing cholangitis, ulcerative colitis, Crohn’s disease.<br />
• Serum dilution 1 : 100; conjugate class anti-human IgG, POD-labelled.<br />
• 6 relevant anti-granulocyte antibodies can be detected simultaneously and<br />
monospecifically: autoantibodies against pro teinase 3, lactoferrin, myeloperoxidase,<br />
elastase, cathepsin G, BPI.<br />
• 1-point calibration, semi-quantitative. Calibrator pool and serum sample on the<br />
same microplate strip.<br />
• Native antigens, purified by affinity chromatography.<br />
• Available individual ELISA (3-point calibration, quantitative):<br />
Antigen Order No.<br />
Proteinase 3 (PR3-hn-hr: native/recombinant) EA 1201-9601-2 G<br />
Myeloperoxidase EA 1211-9601 G
<strong>EUROIMMUN</strong> Medizinische<br />
Labordiagnostika<br />
AG<br />
Autoantibodies against CIBDRelevant Antigens<br />
Indirect Immunofluorescence test: CIBD Profile<br />
• Screening and differentiation test for the detection of antibodies in chronic<br />
inflammatory bowel diseases (CIBD): pancreas antigens rPAg1 and rPAg2,<br />
intestinal goblet cells, granulocytes (EOH), lactoferrin-specific (LFS) granulocytes,<br />
Saccharomyces cerevisiae.<br />
• Indication: Crohn‘s disease, ulcerative colitis.<br />
• Initial dilution: pancreas antigens, intestinal goblet cells and granulocytes 1:10<br />
(IgA, IgG), S. cerevisiae 1:100 (IgA), 1:1000 (IgG).<br />
• For serological diagnosis of ulcerative colitis the indirect immunofluorescence<br />
test uses goblet cells (differentiated intestinal cells) for the detection of<br />
autoantibodies against intestinal goblet cells, and ethanol-fixed (EOH-fixed)<br />
granulocytes for the detection of anti-neutrophil cytoplasmic antibodies<br />
(ANCA). Another important antibody associated with ulcerative colitis is directed<br />
against DNA-bound lactoferrin. For the determination of these autoantibodies<br />
granulocytes selectively reacting with lactoferrin (LFS granulocytes) are used.<br />
• The investigation of antibodies against against exocrine pancreas (rPAg 1 + 2)<br />
and antibodies against S. cerevisiae is used for serological diagnosis of Crohn‘s<br />
disease. For the detection of antibodies against pancreas antigens rPAg1<br />
(CUZD1) and rPAg2 (gP2) transfected cells are used as the standard substrate.<br />
• Differential diagnosis is most efficient using a substrate combination of goblet<br />
cells, granulocytes, rPAg1 / rPAg2 and S. cerevisiae. If LFS granulocytes are<br />
used in addition (e.g. FA 1391-####-4), the hit rate for the serological diagnosis<br />
of chronic inflammatory bowel diseases can be increased significantly.<br />
Transfected cells: autoantibodies against CUZD1<br />
and GP2 (top); EOH-fixed and LFS granulocytes<br />
(middle); intestinal goblet cells and S. cerevisiae<br />
(bottom).<br />
Order No. Formats<br />
FA 1391-####-4 page 127.<br />
— 49 —<br />
<strong>EUROIMMUN</strong> <strong>Product</strong>s for the<br />
Determination of Autoantibodies
<strong>EUROIMMUN</strong> Medizinische<br />
M edizinische<br />
Labordiagnostika<br />
Labordiagnostika<br />
<strong>EUROIMMUN</strong> AG<br />
AG<br />
<strong>EUROIMMUN</strong> <strong>Product</strong>s for the<br />
Determination of Autoantibodies<br />
Cost-Effective Strategy for the Detection of<br />
Autoantibodies against Granulocyte Cytoplasm (ANCA)<br />
Screening test using indirect immunofluorescence BIOCHIP sextet: granulocytes (EOH), granulocytes<br />
(HCHO), EUROPL<strong>US</strong> microdots PR3, MPO and GBM, and human epithelial cells (HEp-2) plus granulocytes<br />
A<br />
Granulocytes<br />
(EOH-fixed)<br />
B<br />
Granulocytes<br />
(HCHO-fixed)<br />
C<br />
EUROPL<strong>US</strong><br />
PR3<br />
microdots<br />
At B cytoplasmic<br />
fluorescence<br />
pANCA & GBM & MPO<br />
ELISA: Anti-MPO/Anti-GBM<br />
AAb against<br />
MPO<br />
Goodpasture Syn. 30-60 %<br />
F<br />
HEp-2 cells<br />
+<br />
granulocytes<br />
(EOH-fixed)<br />
E<br />
EUROPL<strong>US</strong><br />
GBM<br />
microdots<br />
D<br />
EUROPL<strong>US</strong><br />
MPO<br />
microdots<br />
MPA 53 %<br />
AAb against<br />
GBM<br />
Perinuclear fluorescence of<br />
granulocytes (A, F)<br />
MPA 42-70 %<br />
CSS 18-60 %<br />
SLE 9-25 %<br />
RA 3-25 %<br />
pANCA<br />
At B no cytoplasmic<br />
fluorescence<br />
DNA-ANCA<br />
UC 67 %<br />
CD 7 %<br />
PSC 87 %<br />
ELISA: ANCA Profile<br />
AAb against PR3, MPO,<br />
elastase, cathepsin G,<br />
BPI, lactoferrin<br />
WG 5 % (BPI)<br />
MPA 53 % (MPO)<br />
MPA 3 % (LF)<br />
RA, vasculitides 45 % (LF)<br />
SLE 6 % (EL)<br />
Fluorescence of all cell nuclei<br />
(A, F)<br />
only ANA<br />
Cytoplasmic fluorescence<br />
of granulocytes (B)<br />
pANCA & ANA & MPO<br />
MPA 42-70 %<br />
Qualified ANA diagnostics: screening test using<br />
HEp-2 cells + granulocytes (F), differentiation<br />
using ELISA, EUROASSAY, EUROLINE, Westernblot<br />
AAb against dsDNA, ssDNA, nucleosomes,<br />
histones, nuclear membrane, nRNP/Sm, Sm,<br />
SS-A, Ro-52, SS-B, Ku, cyclin I (PCNA), mitosin<br />
(CENP-F, cyclin II), nuclear dots, centromeres<br />
(CENP B), spindle fibres, midbody, centrioles,<br />
Scl-70, PM-Scl, fibrillarin, RNA polymerase I,<br />
NOR, ribosomal P-proteins, Jo-1, PL-7, PL-12,<br />
Mi-2, mitochondria (AMA), lysosomes, Golgi<br />
apparatus, vimentin, tropomyosin, actin.<br />
Fluorescence of all cell nuclei<br />
(A, F), granuloc. accentuated (F)<br />
ANA and pANCA<br />
Nuclei of granuloc. brighter than<br />
nuclei of HEp-2 cells (F), B neg.<br />
DNA-ANCA & ANA<br />
cANCA<br />
WG 80-90 %<br />
MPA 10-15 %<br />
CSS 10-20 %<br />
PAN < 9 %<br />
ELISA: Anti-PR3-hn-hr / ANCA Profile (BPI)<br />
AAb against<br />
PR3-hn-hr<br />
WG 80-90 %<br />
Cytoplasmic fluorescence<br />
of granulocytes (A, B, F)<br />
ANCA reaction at A,<br />
ANA reaction at F<br />
DNA-ANCA? & ANA<br />
AAb against<br />
BPI<br />
WG 5 %<br />
See <strong>EUROIMMUN</strong> poster: " Strategy for Determination<br />
of Autoantibodies against Cell Nuclei (ANA)<br />
and Cytoplasm Components"<br />
The highest diagnostic sensitivity in the determination of autoantibodies against neutrophil granulocytes (ANCA) is achieved by using indirect immunofluorescence<br />
and assays with defined target antigens (particularly PR3, MPO and GBM) simultaneously at the start. However, under the pressure of cost optimisation, an immunofluorescence<br />
test may be performed on its own and then followed up by specific ELISA tests only if the result is positive. Ethanol-fixed human granulocytes<br />
are the standard substrate for indirect immunofluorescence. With this substrate two relevant fluorescence patterns can be differentiated: the cytoplasmic type<br />
(cANCA) associated with Wegener’s granulomatosis and the perinuclear type (pANCA), which indicates a range of various diseases. The differentiation of pANCA<br />
from antibodies against cell nuclei (ANA) is often difficult. Therefore, HEp-2 cells (possibly with sedimented granulocytes) or primate liver are used as an additional<br />
substrate. If ANA and pANCA occur together, the granulocytes show a much brighter fluorescence than the cell nuclei. Thanks to <strong>EUROIMMUN</strong> BIOCHIPs it is not<br />
necessary to incubate human epithelial cells on a second slide in parallel for the exclusion of cell nucleus antibodies, since all substrates are present in one and the<br />
same test field. A third BIOCHIP with formalin-fixed human granulocytes detects a large proportion of the diagnostically relevant antibodies against myeloperoxidase,<br />
whereas other pANCA (which are particularly important in gastroenterology) and almost all antibodies against cell nuclei (whose differentiation is a separate<br />
chapter in autoantibody diagnostics) are generally completely suppressed. The EUROPL<strong>US</strong> substrates PR3, MPO, GBM help to confirm diagnosis and allow a quick<br />
and reliable interpretation of results even in problematic cases.<br />
ANA: anti-nuclear antibodies BPI: bactericidal permeability increasing protein cANCA: anti-neutrophil cytoplasmic antibodies, cytoplasmic type CD: Crohn‘s disease CSS: Churg-Strauss syndrome EL: elastase EOH: ethanol<br />
HCHO: formalin HEp-2: human epithelial cells IFT: immunofluorescence test LF: lactoferrin MPA: microscopic polyangiitis MPO: myeloperoxidase PAN: polyarteritis nodosa pANCA: anti-neutrophil cytoplasmic antibodies,<br />
perinuclear type PR3: proteinase 3 PSC: primary sclerosing cholangitis RA: rheumatoid arthritis SLE: systemic lupus erythematosus UC: ulcerative colitis WG: Wegener‘s granulomatosis<br />
<strong>EUROIMMUN</strong> — 50 — AG · D-23560 Luebeck (Germany) · Seekamp 31 · Phone +49 451 58550 · Fax 5855591 · E-mail euroimmun@euroimmun.de<br />
HA_1200_I_UK_A04, 09/2011
<strong>EUROIMMUN</strong> Medizinische<br />
Labordiagnostika<br />
AG<br />
Autoantibodies against Phospholipase A2 Receptor (PLA2R)<br />
Indirect Immunofluorescence test: Anti-Phospholipase A2 Receptor<br />
• Detection of antibodies against phospholipase A2 receptor (PLA2R).<br />
• Indication: primary membranous glomerulonephritis (MGN, also idiopathic<br />
membranous nephropathy IMN).<br />
• Initial dilution 1 : 10; conjugate class anti-human IgG, FITC labelled.<br />
• The Anti-Phospholipase A2 receptor IIFT uses transfected cells as the standard<br />
substrate. Antibodies against PLA2R react with the transfected cells of the test<br />
substrate. They induce a cytoplasmic fluorescence, with some fluorescence of<br />
the cell membrane.<br />
• The detection of antibodies against phospholipase A2 receptors (PLA2R) is very<br />
specific and helps to differentiate primary membranous nephropathy from the<br />
secondary form of the disease.<br />
Microplate ELISA: Anti-Phospholipase A2 Receptor<br />
• Monospecific<br />
(PLA2R).<br />
detection of antibodies against phospholipase A2 receptor<br />
• Indication: primary membranous glomerulonephritis (MGN, also idiopathic<br />
membranous nephropathy IMN).<br />
• Serum dilution 1: 100, conjugate class anti-human IgG, POD-labelled.<br />
• 5-point calibration, quantitative.<br />
• Antigen: recombinant, human c-DNA expressed in human cell line.<br />
• The anti-PLA2R antibody titer is a marker that indicates whether immunosuppressive<br />
therapy is required and which type should be used. It is also useful<br />
for monitoring the patient during therapy.<br />
Transfected cells: antibodies against PLA2R.<br />
Order No. Formats<br />
FA 1254-####-50 page 125<br />
Incubated ELISA Anti-PLA2R.<br />
Order No. Formats<br />
EA 1254-9601 G page 89<br />
— 51 —<br />
<strong>EUROIMMUN</strong> <strong>Product</strong>s for the<br />
Determination of Autoantibodies
<strong>EUROIMMUN</strong> Medizinische<br />
Labordiagnostika<br />
AG<br />
<strong>EUROIMMUN</strong> <strong>Product</strong>s for the<br />
Determination of Autoantibodies<br />
Primate liver and gliadin (GAF-3X) BIOCHIP: antibodies<br />
against endomysium and gliadin.<br />
Order No. Formats<br />
FA 1914-####-1 A or G page 135<br />
Incubated ELISA Anti-Gliadin (GAF-3X).<br />
Order No. Formats<br />
EV 3011-9601 A or G page 100<br />
Incubated ELISA Anti-Tissue Trans glutaminase<br />
(Endomysium).<br />
Order No. Formats<br />
EA 1910-9601 A or G page 91<br />
— 52 —<br />
Antibodies against Endomysium and gliadin<br />
Indirect Immunofluorescence Test: EUROPL<strong>US</strong> Primate Liver<br />
and Gliadin (GAF-3X) BIOCHIPs<br />
• Detection of antibodies against endomysium and gliadin.<br />
• Indications: gluten-sensitive enteropathy (celiac disease, non-tropical sprue),<br />
Duhring’s herpetiform derma titis.<br />
• Initial dilution 1 : 10; conjugate class anti-human IgA or IgG, primate absorbed,<br />
FITC-labelled.<br />
• Autoantibodies against endomysium react with many types of tissue, e.g.<br />
primate oesophagus. The most suitable substrate, however, is primate liver: in<br />
the case of a positive sample, filamentous linings of the intralobular sinusoids<br />
react.<br />
• With the gliadin (GAF-3X)-coated BIOCHIP, antibodies against gliadin can be<br />
analyzed in one and the same test procedure.<br />
• Both anti-endomysium antibodies and antibodies against gliadin (class IgA) are<br />
reliable serological markers for an active gluten-sensitive entero pathy. Therefore,<br />
their determination can in many cases replace endoscopy and biopsy.<br />
Microplate ELISA: Anti-Gliadin (GAF-3X)<br />
• Monospecific detection of antibodies against gliadin.<br />
• Indications: gluten-sensitive enteropathy (celiac disease, non-tropical sprue),<br />
Duhring’s herpetiform derma titis.<br />
• Serum dilution 1 : 200; conjugate class anti-human IgA or IgG, POD-labelled.<br />
• 3-point calibration. Identical incubation conditions and times: the investigation<br />
of IgA and IgG antibodies can be combined without difficulty on one and the<br />
same microplate.<br />
• Antigen: Gliadin-analogue fusion peptide (GAF-3X).<br />
• The quantitative determination of antibodies against gliadin is very suitable for<br />
monitoring the progress of the disease, compliance with a gluten-free diet, or a<br />
gluten tolerance test.<br />
Microplate ELISA: Anti-Tissue Transglutaminase (Endomysium)<br />
• Monospecific detection of antibodies against tissue transglutaminase.<br />
• Indications: gluten-sensitive enteropathy (celiac disease, non-tropical sprue),<br />
Duhring’s herpetiform derma titis.<br />
• Serum dilution 1 : 200; conjugate class anti-human IgA or IgG, POD-labelled.<br />
• 3-point calibration, quantitative.<br />
• Antigen: recombinant, expression with the baculovirus vector in insect cells.
<strong>EUROIMMUN</strong> Medizinische<br />
Labordiagnostika<br />
AG<br />
<strong>EUROIMMUN</strong> PRODUCTS FOR INFECTIO<strong>US</strong> SEROLOgY<br />
— 53 —<br />
<strong>EUROIMMUN</strong> <strong>Product</strong>s for<br />
Infectious Serology
<strong>EUROIMMUN</strong> Medizinische<br />
Labordiagnostika<br />
AG<br />
<strong>EUROIMMUN</strong> <strong>Product</strong>s for<br />
Infectious Serology<br />
Antibodies against Borrelia afzelii, VlsE (top),<br />
Borrelia burgdorferi and OspC (bottom).<br />
Order No. Formats<br />
FI 2136-####-1 G or M page 137.<br />
Incubated ELISA Anti-Borrelia plus VlsE.<br />
Order No. Formats<br />
EI 2132-9601 G or M page 94<br />
Table-based evaluation of the CSQ rel.<br />
Order No. Formats<br />
EI 2132-9601-L G or M page 94<br />
— 54 —<br />
Antibodies against Borrelia<br />
Indirect Immunofluorescence Test: EUROPL<strong>US</strong> Anti-Borrelia<br />
afzelii, Borrelia burgdorferi, VlsE and OspC Antigen<br />
• Sensitive screening test for the detection of anti-Borrelia antibodies.<br />
• Indications: erythema chronicum migrans, lymphadenosis cutis benigna, lymphocytic<br />
meningoradiculitis, carditis, arthritis, acrodermatitis chronica atrophicans,<br />
neuroborreliosis.<br />
• Initial dilution 1 : 10 (IgM), 1 : 100 (IgG).<br />
• If antibodies against Borrelia afzelii or Borrelia burgdorferi are present, a distinct<br />
fluorescence of the bacteria in the smear is obtained.<br />
• With the VlsE or OspC coated BIOCHIPs antibodies against the highly specific<br />
and highly sensitive marker antigens VlsE (IgG) or OspC (IgM) can be determined<br />
monospecifically in one and the same test procedure. If these antigens fluoresce<br />
the antibody result is positive even if the bacteria smears show a negative<br />
reaction. Thus, the BIOCHIP Mosaic helps to increase specificity and sensitivity<br />
in Borrelia diagnostics.<br />
• The BIOCHIP can be supplemented as required using further substrates, e.g.<br />
Borrelia burgdorferi sensu stricto (strains CH or <strong>US</strong>A) and TBE virus infected<br />
cells.<br />
Microplate ELISA: Anti-Borrelia plus VlsE<br />
• Sensitive screening test for the detection of anti-Borrelia antibodies.<br />
• Indications: erythema chronicum migrans, lymphadenosis cutis benigna, lymphocytic<br />
meningoradiculitis, carditis, arthritis, acrodermatitis chronica atrophicans,<br />
neuroborreliosis.<br />
• Serum dilution 1:101; conjugate class anti-IgG or anti-IgM (VlsE: -IgG only), PODlabelled.<br />
• 3-point calibration (IgG and IgM). Identical incubation conditions and times: all<br />
tests can be combined without difficulty on one and the same microplate.<br />
• Antigens: extract of Borrelia burgdorferi sensu stricto, Borrelia garinii and<br />
Borrelia afzelii (whole antigen) / recombinant VlsE from Borrelia burgdorferi<br />
sensu stricto (IgG). VlsE (variable major protein-like sequence, expressed) is<br />
a surface protein of Borrelia which is expressed exclusively in vivo and which<br />
contains conserved and highly immunogenic epitopes.<br />
• IgM test kit (Anti-Borrelia) includes IgG/RF absorbent in sample buffer for IgG<br />
absorption in preparation for the determination of specific IgM class antibodies.<br />
Microplate ELISA: Anti-Borrelia plus VlsE, Antibody Determination<br />
in Serum and Cerebrospinal Fluid for Detection of Intrathecal<br />
Synthesis of Specific Antibodies against Borrelia<br />
• Antibody determination in serum and cerebrospinal fluid (CSF).<br />
• Indication: Neuroborreliosis.<br />
• CSF dilution 1 : 2, serum dilution 1 : 404; conjugate class anti-IgG or anti-IgM,<br />
POD-labelled.<br />
• 4-point calibration, quantitative.
<strong>EUROIMMUN</strong> Medizinische<br />
Labordiagnostika<br />
AG<br />
Anti-Borrelia EUROLINE-RN-AT<br />
Antibodies against Borrelia<br />
• Specific confirmatory test for the detection of antibodies against Borrelia.<br />
• Indications: erythema chronicum migrans, lymphadenosis cutis benigna, lymphocytic<br />
meningoradiculitis, carditis, arthritis, acrodermatitis chronica atrophicans,<br />
neuroborreliosis.<br />
• The Anti-Borrelia EUROLINE provides a unique mixture of Borrelia specific<br />
antigens in a user-friendly line blot format. In addition to the major serological<br />
early markers OspC (IgM) and VlsE (IgG) – in each case from B. afzelii, B.<br />
burgdorferi and B. garinii – it contains highly specific p39 (Bmp) and the late<br />
marker p38. Five new, recombinant designer antigens (p18, p19, p20, p21, p58)<br />
having a very high specificity (IgG) were identified using bioinformatic analysis<br />
of the Borrelia genome. For the first time, lipids which have been proven to be<br />
immunoreactive and were extracted from the Borrelia membrane are available<br />
on the line blot.<br />
• The Anti-Borrelia EUROLINE-RN-AT (IgG) can also be used for qualitative detection<br />
of specific intrathecal antibody synthesis in neuroborreliosis.<br />
• The serological hit rate is increased by 10% by using three OspC variants in the<br />
IgM test.<br />
• Serum dilution 1 : 51; conjugate class anti-IgG or anti-IgM, AP-labelled.<br />
EUROLINE-WB: Anti-Borrelia (Whole Antigen plus VlsE)<br />
• Specific confirmatory test for the detection of antibodies against Borrelia.<br />
• EUROLINE-WB is a combination of westernblot and line blot techniques. An SDS<br />
extract of a Borrelia afzelii strain is electrophoretically separated according to<br />
molecular mass and transferred onto a nitrocellulose membrane. A membrane<br />
chip coated with highly purified recombinant VlsE-Antigen is then added to the<br />
westernblot strips.<br />
• By additionally determining antibodies against VlsE the serological hit rate<br />
can be increased by 20% compared to whole extract Western blots and by 30%<br />
compared to recombinant antigen Westernblots. Of all recombi nant antigens,<br />
VlsE possesses the highest sensitivity for the detection of a Borrelia infection.<br />
Over 85% of IgG-positive sera could be identified at a glance by assessing the<br />
VlsE band. VlsE allows detection of antibodies against all Borrelia species, and the<br />
risk of a false negative reaction due to species differences is ten times lower.<br />
• Serum dilution 1 : 51; conjugate class anti-IgG or anti-IgM, AP-labelled.<br />
VlsE B. burgd.<br />
p41<br />
p39<br />
OspC B. afzelii<br />
OspC B. burgd.<br />
OspC B. garinii<br />
IgM<br />
Control band<br />
Automated Evaluation of Incubated Membrane Strips with the System EUROLineScan<br />
VlsE B. afzelii<br />
VlsE B. burgdorferi<br />
VlsE B. garinii<br />
Lipid B. afzelii<br />
Lipid B. burgdorferi<br />
p83<br />
p41<br />
p39<br />
VlsE<br />
p 83<br />
p 41, Flag.<br />
p 39, Bmp A<br />
p 31, Osp A<br />
p 30<br />
p 25, Osp C<br />
• The program EUROLineScan from <strong>EUROIMMUN</strong> has been developed to enable quantitative evaluation of membrane based test systems,<br />
facilitate management of data, and provide detailed documentation of results — tasks which have until now required con siderable time.<br />
• First, the incubated test strips are scanned using a flatbed scanner or camera system.<br />
• EUROLineScan recognizes the position of the strips, even if they have been placed inexactly, identifies the bands, and measures their<br />
inten sity. The auto mated evalu ation can be moni tored, and it is possible to supplement the data manually.<br />
• The results are then saved together with the image data. It is no longer necessary to archive (potentially infectious) incubated test strips.<br />
A separate results sheet can be produced for each patient.<br />
p 21<br />
p 19<br />
p 17.<br />
OspC<br />
p58<br />
p21<br />
p20<br />
p19<br />
p18<br />
Igg<br />
Incubated Anti-Borrelia EUROLINE-RN-AT .<br />
Order No. Formats<br />
DN 2131-3201 G or M page 83<br />
Control band<br />
Control band<br />
Incubated EUROLINE-WB Anti-Borrelia.<br />
Order No. Formats<br />
DY 2131-1601-1 G or M page 87.<br />
— 55 —<br />
<strong>EUROIMMUN</strong> <strong>Product</strong>s for<br />
Infectious Serology
<strong>EUROIMMUN</strong> Medizinische<br />
Labordiagnostika<br />
AG<br />
<strong>EUROIMMUN</strong> <strong>Product</strong>s for<br />
Infectious Serology<br />
— 56 —<br />
Antibodies against EpsteinBarr virus (EBv)<br />
Patient 1: EBV-CA (IgG) EBV-CA (IgG): urea-treated EBV-CA (IgM) EBV-EA (IgG) EBNA (IgG)<br />
Patient 2: EBV-CA (IgG) EBV-CA (IgG): urea-treated EBV-CA (IgM) EBV-EA (IgG) EBNA (IgG)<br />
Indirect Immunofluorescence Test: BIOCHIP Sequence EBV<br />
• Gold standard for the determination of antibodies against the EBV-CA antigens (Epstein-Barr virus capsid antigen), EBV-EA (Epstein-<br />
Barr early antigen) and EBNA (Epstein-Barr nuclear antigen).<br />
• Indication: infectious mononucleosis, Burkitt‘s lymphoma, nasopharyngeal carcinoma (NPC).<br />
• IgG antibodies against EBV-CA indicate an EBV infection. An at least twofold increase in titer and the absence of antibodies against<br />
EBNA at the same time is characteristic for the early phase of the infection. IgM antibodies against EBV-CA and antibodies against<br />
EBV-EA can also be found in acute infections, but they do not necessarily always occur. The presence of antibodies against EBNA<br />
generally indicates the late phase of an EBNA infection.<br />
• In cases of an acute EBV infection which cannot be reliably discriminated from a relapse or reinfection, the determination of the<br />
antibody avidity using a modified immunofluorescence test as an additional parameter is useful. This requires an additional incubation<br />
with urea solution (ZF 1130-0801). The determination of low-avidity antibodies against EBV-CA indicates an acute infection.<br />
• For the monospecific confirmation of EBV-CA antibodies in the same test procedure the BIOCHIP containing ECV-CA is supplemented<br />
with the antigen substrates gp125 antigen (native) and p19 antigen (recombinant) (EUROPL<strong>US</strong> FI 27.91-####-20 G or M).<br />
• For highly differentiated diagnostics the BIOCHIP Sequence EBV (FI 2799-####-1 X) can be supplemented by using the antigens gp125<br />
and p19 (EUROPL<strong>US</strong> FI 2799-####-21 X).<br />
• Due to the high prevalence of anti-EBV-CA IgA in NPC patients, the parameter is well suited for screening. Confirmation of the result<br />
by determination of IgA antibodies against EBV-EA is recommended. Further anti-EBV test kits for indirect immunofluorescence:<br />
Substrate Order No. Substrate Order No.<br />
Order No. Formats<br />
FI 2799-####-21 X page 144<br />
Incubated ELISA Anti-EBNA-1.<br />
Order No. (anti-EBNA-1) Formats<br />
EI 27.93-9601 G page 100<br />
EBV-CA FI 27.91-#### A, G or M EBV-EA FI 27.95-#### A or G<br />
EBNA FI 27.93-#### G EBV-CA & EBV-EA FI 27.91-####-2 A or G<br />
Microplate ELISA: Anti-EBV-CA, Anti-EBNA-1, Anti-EBV-EA<br />
• Specific confirmatory test for antibodies against EBV-CA, EBNA-1 or EBV-EA.<br />
• Indications: infectious mononucleosis, Burkitt’s lymphoma, nasopharyngeal<br />
carcinoma.<br />
• Serum dilution 1 : 101; conjugate class anti-IgA, -IgG or IgM, POD-labelled.<br />
• 1-point calibration, semi-quantitative (IgA, IgM) or 3-point calibration, quantitative<br />
(IgG). Identical incubation conditions and times: all tests can be combined<br />
without difficulty on one and the same microplate.<br />
• EBV-CA: native antigen, purified by affinity chromatography; EBNA-1 and EBV-EA:<br />
recom binant antigen.<br />
• IgM test kit includes IgG/RF absorbent in sample buffer for IgG absorption in<br />
preparation for the determination of specific IgM class antibodies.<br />
• Available individual ELISA:<br />
Antigen Order No.<br />
EBV-CA EI 27.91-9601 A, G or M<br />
EBNA-1 EI 27.93-9601 G<br />
EBV-EA EI 27.95-9601 A, G or M<br />
fresh infection<br />
previous infection
<strong>EUROIMMUN</strong> Medizinische<br />
Labordiagnostika<br />
AG<br />
Antibodies against EpsteinBarr virus (EBv)<br />
Determination of Low-Avidity Antibodies against EBV-CA<br />
• An alternative principle for the serological diagnosis of fresh infections with EBV<br />
has been established by investigating the antibody avidity.<br />
• The first reaction of the immune system following an infection is the formation<br />
of low-avidity antibodies. As the infection proceeds, increasingly antigen-adapted<br />
IgG is formed, and avidity grows. As long as high-avidity IgG is not yet detected<br />
in the serum, it can be assumed that the infection is still in an early stage.<br />
• To identify low-avidity antibodies against EBV-CA in a patient’s serum, two<br />
micro plate ELISA or immunofluorescence tests are performed in parallel: one<br />
test is carried out in the conventional way, the other one includes urea treatment<br />
between incubations with patient’s serum and per oxidase-labelled anti-human<br />
IgG, resulting in the detachment of low-avidity antibodies from the antigens.<br />
• Low-avidity antibodies against EBV-CA are present if the ELISA extinction is<br />
significantly reduced by urea treatment. For an objective interpretation, the<br />
relative avidity index (RAI) can be calculated out of the measured values with<br />
and without urea incubation.<br />
• With the immunofluorescence test the presence of low-avidity antibodies has<br />
been proved if the test using urea treatment gives a far weaker fluorescence<br />
than the two-step test.<br />
EUROLINE: EBV Profile 2<br />
• Differentiation of antibodies against Epstein-Barr virus antigens.<br />
• Indications: infectious mononucleosis, Burkitt’s lymphoma, nasopharyngeal<br />
carcinoma.<br />
• Serum dilution 1 : 51; conjugate class anti-human IgG or IgM, AP-labelled.<br />
• With the EUROLINE EBV Profile 2, five different antibodies can be determined:<br />
anti bodies against VCA gp125, VCA p19, EBNA-1, p22, EA-D.<br />
• Recombinant antigens (exception: VCA gp125, native, purified by affinity chromatography).<br />
• EBNA-1 (IgG) negative und VCA (IgG and IgM) positive: acute (fresh) infection.<br />
• EBNA-1 (IgG) and VCA (IgG) positive and VCA (IgM) negative: late phase of<br />
infection.<br />
• EBNA-1 (IgG) negative, but band p22 (IgG) and VCA (IgG) positive: late phase of<br />
infection with loss of anti-EBNA-1.<br />
• Test strips can be automatically incubated and evaluated using the systems<br />
EUROBlotMaster und EUROLineScan (see page 27.).<br />
Westernblot: Anti-EBV<br />
• Specific confirmatory test for the detection of antibodies against Epstein-Barr<br />
virus antigens, differentiation of antibodies against Epstein-Barr virus antigens.<br />
• Indications: infectious mononucleosis, Burkitt’s lymphoma, nasopharyngeal<br />
carcinoma.<br />
• Serum dilution 1 : 51; conjugate class anti-IgG or anti-IgM, AP-labelled.<br />
• Antigens: whole antigen, SDS extract.<br />
• Bands from all specific antigens are included and clearly separated.<br />
• EBNA-1 (IgG) negative und VCA (IgG and IgM) positive: acute (fresh) infection.<br />
• EBNA-1 (IgG) and VCA (IgG) positive and VCA (IgM) negative: late phase of<br />
infection.<br />
• EBNA-1 (IgG) negative, but band p22 (IgG) and VCA (IgG) positive: late phase of<br />
infection with loss of anti-EBNA-1.<br />
• Test strips can be automatically incubated and evaluated using the systems<br />
EUROBlotMaster und EUROLineScan (see page 27.).<br />
Number of Patients<br />
10<br />
8<br />
6<br />
4<br />
2<br />
0<br />
0 25 50 75 100<br />
VCA gp125<br />
VCA p19<br />
EBNA-1<br />
p22<br />
EA-D<br />
Order No. Formats<br />
DN 27.90-1601-2 G or M page 83<br />
VCA 125<br />
VCA 65<br />
VCA 40/41/42<br />
Relative Avidity Index (RAI) in %<br />
Avidity of antibodies against EBV-CA (IgG).<br />
VCA 33<br />
VCA 22<br />
RAI = E with urea<br />
E without urea<br />
Incubated Westernblot Anti-EBV.<br />
Order No. Formats<br />
DY 27.90-1501 G or M page 88<br />
Primary Infection<br />
Previous Infection<br />
Order No. (IIFT)<br />
FI 2791-#### X<br />
Order No. (ELISA) Formats<br />
EI 27.91-####-1 G page 99 and 143<br />
Control IgG/IgM<br />
Incubated EUROLINE EBV Profile 2.<br />
EA R<br />
EBNA-1<br />
EA D<br />
Control band<br />
— 57. —<br />
<strong>EUROIMMUN</strong> <strong>Product</strong>s for<br />
Infectious Serology
<strong>EUROIMMUN</strong> Medizinische<br />
Labordiagnostika<br />
AG<br />
<strong>EUROIMMUN</strong> <strong>Product</strong>s for<br />
Infectious Serology<br />
Antibodies against Helicobacter pylori.<br />
Order No. Formats<br />
FI 2080-#### A, G or M page 137.<br />
Incubated ELISA Anti-Helicobacter pylori.<br />
Order No. Formats<br />
EI 2080-9601 A or G page 94<br />
— 58 —<br />
Antibodies against helicobacter pylori<br />
p 120, CagA<br />
p 95, VacA<br />
p 66, UreB<br />
p 33<br />
p 30<br />
p 29, UreA<br />
p 26<br />
p 19, OMP<br />
p 17.<br />
Alignment bar<br />
Control band<br />
Incubated EUROLINE-WB Anti-Helicobacter pylori.<br />
Order No. Formats<br />
DY 2080-1601-1 A or G page 87.<br />
Indirect Immunofluorescence Test: Anti-Helico bacter pylori<br />
• Sensitive screening test for the detection of antibodies against Helicobacter<br />
pylori.<br />
• Indications: gastritis, ulcus ventriculi et duodeni. Late consequences: MALT<br />
lympho mas and adenocarcinomas.<br />
• Initial dilution 1 : 10 (IgM), 1 : 100 (IgG), 1 : 32 (IgA).<br />
• If antibodies against Helicobacter pylori are present, a distinct fluorescence of<br />
the bacteria in the smear is obtained.<br />
• A positive IgA result correlates well with the activity of a gastritis. An increased<br />
IgG antibody titer is considered to be a marker for chronic infections. A significant<br />
drop in the IgG antibody titer about 6 months after therapy is a sign of<br />
success.<br />
• The BIOCHIP can be supplemented as required with further substrates, e.g. other<br />
infectious agents or tissue sections of primate stomach.<br />
Microplate ELISA: Anti-Helicobacter pylori<br />
• Sensitive screening test for the detection of antibodies against Helicobacter<br />
pylori.<br />
• Indications: gastritis, ulcus ventriculi et duodeni. Late consequences: MALT<br />
lympho mas and adenocarcinomas.<br />
• Serum dilution 1 : 101; conjugate class anti-IgA or anti-IgG, POD-labelled.<br />
• Antibodies against Helicobacter pylori antigens can be determined quantitatively<br />
in RU/ml.<br />
• 1-point calibration, semi-quantitative (IgA) or 3-point calibration, quantitative<br />
(IgG). Identical incubation conditions and times: both tests can be combined<br />
without difficulty on one and the same microplate.<br />
• Native antigens.<br />
EUROLINE-WB: Anti-Helicobacter pylori<br />
• Specific confirmatory test for the detection of antibodies against Helicobacter<br />
pylori.<br />
• Indications: gastritis, ulcus ventriculi et duodeni. Late consequences: MALT<br />
lympho mas and adenocarcinomas.<br />
• Serum dilution 1 : 50; conjugate class anti-IgA or anti-IgG, AP-labelled.<br />
• EUROLINE-WB is a combination of westernblot and line blot techniques. An<br />
SDS extract of a Helicobacter pylori strain is electrophoretically separated<br />
according to molecular mass and transferred onto a nitrocellulose membrane<br />
(westernblot). Two membrane chips coated with highly purified recombinant<br />
CagA and VacA are subsequently applied to the westernblot strips.<br />
• Bands from all specific antigens are included and clearly separated.<br />
• Test strips can be automatically incubated and evaluated using the systems<br />
EUROBlotMaster und EUROLineScan (see page 27.).
<strong>EUROIMMUN</strong> Medizinische<br />
Labordiagnostika<br />
AG<br />
Antibodies against herpes Simplex virus (hSv)<br />
Indirect Immunofluorescence Test: BIOCHIP Mosaic HSV-1/HSV-2<br />
• Sensitive screening test for the detection of antibodies against herpes simplex<br />
viruses.<br />
• Indication: herpes simplex.<br />
• Initial dilution 1 : 10 (IgM), 1 : 100 (IgG).<br />
• If antibodies against herpes simplex-2 virus are present in the sample, a typical<br />
fluorescence of the infected cells is obtained – mainly in the outspread cytoplasm,<br />
less in the cell nuclei.<br />
• As HSV-1 and HSV-2 are morphologically and immunologically closely related,<br />
cross-reactions can occur. Differentiation may be attempted by testing a serum<br />
against both antigen substrates and comparing the titers.<br />
• The BIOCHIP Mosaic can be supplemented as required with further substrates,<br />
e.g. other infectious agents.<br />
• Anti-HSV individual tests for indirect immunofluorescence:<br />
Substrate Order No.<br />
HSV-1 FI 2531-#### G or M<br />
HSV-2 FI 2532-#### G or M<br />
HSV-1 and -2 FI 2531-####-1 G or M<br />
Microplate ELISA: Anti-HSV-1, Anti-HSV-2<br />
• Specific confirmatory tests for antibodies against HSV-1 or HSV-2.<br />
• Indication: herpes simplex.<br />
• Serum dilution 1 : 101; conjugate class anti-IgG or anti-IgM, POD-labelled.<br />
• 1-point calibration, semiquantitative (IgM) or 3-point calibration, quantitative<br />
(IgG). Identical incubation conditions and times: all tests can be combined<br />
without difficulty on one and the same microplate.<br />
• Antigens: type-specific glycoproteins C1 or G2, purified by affinity chromatography.<br />
Cross-reactions do not occur.<br />
• IgM test kit includes IgG/RF absorbent in sample buffer for IgG absorption in<br />
preparation for the determination of specific IgM class antibodies.<br />
• Available individual ELISA:<br />
Antigen Order No.<br />
HSV-1 EI 2531-9601-2 G or M<br />
HSV-2 EI 2532-9601-2 G or M<br />
HSV-1/2-Pool EI 2531-9601-1 G or M<br />
EUROLINE-WB: Anti-HSV<br />
• Specific confirmatory test for the differentiation of antibodies against HSV-1 and<br />
HSV-2.<br />
• Indication: herpes simplex.<br />
• Serum dilution 1 : 51; conjugate class anti-IgG or anti-IgM, AP-labelled.<br />
• EUROLINE-WB is a combination of westernblot and line blot techniques. Proteins<br />
from an SDS extract of HSV-1 are electrophoretically separated according to<br />
molecular mass and transferred onto a nitrocellulose membrane. A membrane<br />
chip coated with HSV-2 type-specific glycoprotein G2 (gG 2), purified by affinity<br />
chromatography, is then added to the westernblot strips.<br />
• Bands from all specific antigens are included and clearly separated.<br />
• The gG 2 band allows the simple differentiation between HSV-1 and HSV-2<br />
infections.<br />
• Test strips can be automatically incubated and evaluated using the systems<br />
EUROBlotMaster und EUROLineScan (see page 27.).<br />
Antibodies against HSV-1 and HSV-2.<br />
Order No. Formats<br />
FI 2531-####-1 G or M page 140<br />
Incubated ELISA Anti-HSV-1.<br />
Order No. (anti-HSV-1) Formats<br />
EI 2531-9601-2 G or M page 96<br />
gC 1<br />
gG 1<br />
gG 2<br />
Alignment bar<br />
Control band<br />
Incubated EUROLINE-WB Anti-HSV.<br />
Order No. Formats<br />
DY 2531-1601-1 G or M page 88<br />
— 59 —<br />
<strong>EUROIMMUN</strong> <strong>Product</strong>s for<br />
Infectious Serology
<strong>EUROIMMUN</strong> Medizinische<br />
Labordiagnostika<br />
AG<br />
<strong>EUROIMMUN</strong> <strong>Product</strong>s for<br />
Infectious Serology<br />
Anti-Chlamydia MIF.<br />
Order No. Formats<br />
FI 2191-####-3 A, G or M page 135<br />
Incubated ELISA Anti-Chlamydia trachomatis.<br />
Order No. Formats<br />
EI 2191-9601 A, G or M page 95<br />
Incubated ELISA Anti-Chlamydia pneumoniae.<br />
Order No. Formats<br />
EI 2192-9601 A, G or M page 95<br />
— 60 —<br />
Antibodies against Chlamydia<br />
Indirect Immunofluorescence test: Anti-Chlamydia MIF (Micro-<br />
Immunofluorescence test)<br />
• Serological gold standard for the determination of antibodies against Chlamydia.<br />
• Indication: trachoma, urogenital infections, lymphogranuloma venereum, laryngitis,<br />
sinusitis, bronchitis, pneumonia, psittacosis.<br />
• Initial dilution 1 : 10 (IgA), 1 : 100 (IgG), 1 : 10 (IgM).<br />
• The micro-immunofluorescence test uses purified elementary bodies of the<br />
species C. trachomatis, C. pneumoniae and C. psittaci as the antigen. The mutual<br />
lipopolysaccharide (LPS) antigen is inactivated to minimise cross reactivity.<br />
• The evaluation of the MIF could be significantly facilitated compared to conventional<br />
test systems by using a cell-based substrate.<br />
• The fourth BIOCHIP with non-infected cells allows a reliable differentiation<br />
between unspecific and specific fluorescence.<br />
Substrate Order No.<br />
Chlamydia trachomatis FI 2191-####-80 A, G or M<br />
Chlamydia pneumoniae FI 2192-####-80 A, G or M<br />
Chlamydia psittaci FI 2193-####-80 A, G or M<br />
Microplate ELISA: Anti-Chlamydia trachomatis<br />
• Monospecific detection of antibodies against Chlamydia trachomatis.<br />
• Indication: trachoma, conjunctivitis, urogenital infections, pneumonia in infants,<br />
lymphogranuloma venereum.<br />
• Serum dilution 1:101, conjugate class anti-human IgA, IgG or IgM, POD-labelled.<br />
• 1-point calibration, semiquantitative (IgA and IgM) or 3-point calibration, quantitative<br />
(IgG).<br />
• Antigen: native MOMP antigen (major outer membrane protein). MOMP is a<br />
transmembrane protein and represents the main part of the outer membrane of<br />
the elementary bodies. Protein purification starts with BGM cells infected with<br />
Chlamydia trachomatis of the serotype K.<br />
• IgM test kit includes IgG/RF absorbent in sample buffer for IgG absorption in<br />
preparation for the determination of specific IgM class antibodies.<br />
Microplate ELISA: Anti-Chlamydia pneumoniae<br />
• Monospecific detection of antibodies against Chlamydia pneumoniae.<br />
• Indication: laryngitis, sinusitis, bronchitis, pneumonia.<br />
• Serum dilution 1: 101, conjugate class anti-human IgA, IgG or IgM, POD-labelled.<br />
• 1-point calibration, semiquantitative (IgA and IgM) or 3-point calibration, quantitative<br />
(IgG).<br />
• Antigen: cell lysate from HL cells, strain CDC/CWL-029.<br />
• IgM test kit includes IgG/RF absorbent in sample buffer for IgG absorption in<br />
preparation for the determination of specific IgM class antibodies.
<strong>EUROIMMUN</strong> Medizinische<br />
Labordiagnostika<br />
AG<br />
Indirect Immunofluorescence Test<br />
Antibodies against Emerging viruses<br />
• Over the past years it has been observed that a number of new viruses ( „ emerging“<br />
or „ reemerging viruses“) and other pathogens has spread worldwide, introducing<br />
since then unknown diseases into previously unaffected regions.<br />
• <strong>EUROIMMUN</strong> offers a broad spectrum of indirect immunofluorescence tests for<br />
the detection of specific antibodies against:<br />
• Many of these substrates are available as single substrate with non-infected<br />
cells or as useful combinations (syndrome or geographically orientated) for the<br />
in vestigation of serum samples.<br />
• IgG absorption as preparatory step for the determination of specific antibodies<br />
of class IgM: page 64.<br />
• Cross reactions within the virus family, especially with Flaviviruses, should be<br />
taken into consideration since they may cause false-positive results. The infectious<br />
agent can be determined by titration of the sample and comparison of<br />
titers.<br />
Microplate ELISA<br />
Pathogen Disease, syndromes see page<br />
Corona virus SARS corona virus (SARS-CoV)<br />
Flaviviruses<br />
Bunyaviruses<br />
Togaviruses<br />
• Monospecific determination of antibodies.<br />
• Serum dilution 1: 101, conjugate class anti-human IgG or IgM, POD-labelled.<br />
• 1-point calibration, semiquantitative (IgM) or 3-point calibration, quantitative<br />
(IgG). Similar incubation conditions and times: All tests can be combined on one<br />
and the same microplate.<br />
• Available single ELISA:<br />
Antigen Order No.<br />
Severe acute respiratory syndrome<br />
(SARS)<br />
TBE virus (TBEV) Tick-borne encephalitis 138<br />
West Nile virus (WNV) West Nile fever, encephalitis 138<br />
Japanese encephalitis virus (JEV) Japanese encephalitis 138, 141<br />
Yellow fever virus (YFV)<br />
Yellow fever, hepatitis, haemorrhagic<br />
fever, arthritis<br />
Dengue virus (DENV, types 1-4) Dengue fever, haemorrhagic fever 138, 141<br />
Hantavirus (types HTNV, PUUV,<br />
SEOV, SAAV, DOBV, SNV, ANDV)<br />
Sandfly fever virus (types SFSV,<br />
SFNV, TOSV, CYPV)<br />
Rift valley fever virus (RVFV)<br />
Crimean-Congo fever virus<br />
(CCHFV-GPC and -N)<br />
HFRS (haemorrhagic fever with renal<br />
syndrome); HCPS (Hantaviral cardiopulmonary<br />
syndrome)<br />
Pappataci fever, meningitis,<br />
encephalitis<br />
Rift valley fever, haemorrhagic fever,<br />
hepatitis<br />
TBE EI 2661-9601 G or M, avidity, Ab determination in CSF<br />
TBE „Vienna“ EI 2661-9601-9 G<br />
WNV EI 2662-9601 G or M, avidity<br />
JEV EI 2663-9601 G or M<br />
Usutu EI 2667.-9601 G<br />
Dengue EI 266b-9601 G or M<br />
Chikungunya EI 293a-9601 G or M<br />
137.<br />
138<br />
139<br />
139<br />
Crimean-Congo haemorrhagic fever 141<br />
139, 141<br />
Chikungunya virus (CHIKV) Chikungunya fever, arthritis 141, 142<br />
Sindbis virus (SINV) Sindbis fever 142<br />
Kinetoplastida Leishmania donovani Visceral leishmaniasis 136<br />
haemosporina<br />
Plasmodium falciparum Malaria tropica 136, 141<br />
Plasmodium vivax Malaria tertiana 136, 141<br />
SARS-CoV TBEV<br />
WNV<br />
JEV YFV DENV<br />
PUUV<br />
ANDV<br />
SFSV<br />
RVFV CCHFV CHIKV<br />
SINV<br />
Leish. donov.<br />
Plasmodium<br />
Incubated ELISA Anti-TBE virus, Anti-WNV, Anti-<br />
Dengue virus.<br />
Order No. (Anti-TBE) Formats<br />
EI 2661-9601 G or M page 98<br />
— 61 —<br />
<strong>EUROIMMUN</strong> <strong>Product</strong>s for<br />
Infectious Serology
<strong>EUROIMMUN</strong> Medizinische<br />
Labordiagnostika<br />
AG<br />
<strong>EUROIMMUN</strong> <strong>Product</strong>s for<br />
Infectious Serology<br />
FI 282110011 g FI 282210011 g<br />
RESPIRATORY TRACT PROFILE 1 EXANThEMA PROFILE 1<br />
Igg IgM Igg IgM<br />
Field A 1 : 10 1 : 10 Field A 1 : 10 1 : 10<br />
V Verification BIOCHIP*** V Verification BIOCHIP***<br />
1. RSV 1. HHV-6<br />
2. Adenovirus type 3 2. Rubella virus*<br />
3. Influenza virus type A (H1N1) 3. Measles virus<br />
4. Influenza virus type A (H3N2) 4. Mumps virus<br />
Field B 1 : 10 1 : 10 Field B 1 : 10 1 : 10<br />
5. Influenza virus type B 5. VZV<br />
6. Parainfluenza virus type 1 6. EBV-CA**<br />
7.. Parainfluenza virus type 2 7.. EBV-EA<br />
8. Parainfluenza virus type 3 8. Treponema pallidum<br />
Field C 1 : 10 1 : 10 Field C 1 : 100 1 : 10<br />
9. Parainfluenza virus type 4 9. HSV-1<br />
10. Bordetella pertussis** 10. HSV-2<br />
11. Bordetella parapertussis** 11. Coxsackie virus type B1<br />
12. Mycoplasma pneumoniae 12. Coxsackie virus type A9<br />
Field D 1 : 100 1 : 10 Field D 1 : 100 1 : 10<br />
13. Coxsackie virus type B1 13. Echo virus type 7.<br />
14. Coxsackie virus type A7. 14. Borrelia afzelii<br />
15. Echo virus type 7. 15. Borrelia burgdorferi sensu stricto (CH)<br />
16. Chlamydia pneumoniae 16. Borrelia garinii<br />
Field E 1 : 100 1 : 100 Field E 1 : 100 1 : 100<br />
17.. Haemophilus influenzae* , ** 17.. CMV<br />
18. Klebsiella pneumoniae* 18. Candida albicans**<br />
19. Legionella pneumophila serotype 1* , ** 19. Candida krusei* , **<br />
20. Legionella pneumophila serotype 12* , ** 20. Candida tropicalis* , **<br />
FI 282310011 g FI 282410011 g<br />
LYMPhADENITIS PROFILE 1 CENTRAL NERvO<strong>US</strong> SYSTEM PROFILE 1<br />
Igg IgM Igg IgM<br />
Field A 1 : 10 1 : 10 Field A 1 : 10 1 : 10<br />
V Verification BIOCHIP*** V Verification BIOCHIP***<br />
1. HIV-1* 1. Rubella virus*<br />
2. HIV-2* 2. Measles virus<br />
3. HHV-6 3. Mumps virus<br />
4. Rubella virus* 4. VZV<br />
Field B 1 : 10 1 : 10 Field B 1 : 10 1 : 10<br />
5. Measles virus 5. Adenovirus type 3<br />
6. Mumps virus 6. EBV-CA**<br />
7.. Adenovirus type 3 7.. Treponema pallidum<br />
8. Parainfluenza virus type 1 8. Toxoplasma gondii**<br />
Field C 1 : 10 1 : 10 Field C 1 : 100 1 : 10<br />
9. EBV-CA** 9. HSV-1<br />
10. EBV-EA 10. HSV-2<br />
11. Toxoplasma gondii** 11. Coxsackie virus type B1<br />
12. Treponema pallidum 12. Coxsackie virus type A7.<br />
Field D 1 : 100 1 : 10 Field D 1 : 100 1 : 10<br />
13. HSV-1 13. Echo virus type 7.<br />
14. HSV-2 14. Borrelia afzelii<br />
15. CMV** 15. Borrelia burgdorferi sensu stricto (CH)<br />
16. Coxsackie virus type B5 16. Borrelia garinii<br />
Field E 1 : 100 1 : 10 Field E 1 : 100 1 : 100<br />
17.. Coxsackie virus type A9 17.. CMV<br />
18. Bartonella henselae** 18. Haemophilus influenzae* , **<br />
19. Chlamydia trachomatis** 19. Listeria monocytogenes 1/2a<br />
20. Chlamydia pneumoniae 20. Listeria monocytogenes 4b<br />
— 62 —<br />
BIOChIP Mosaics for Infectious Serology<br />
(For formats see pages 144/145)<br />
* For clinical evaluation the results must be confirmed with a CE approved test.<br />
** For practical reasons the recommended incubation differs from the standard incubation for this substrate.<br />
*** Field A contains a verification BIOCHIP. The incubation was performed correctly if the dots show a visible colour reaction.<br />
BIOCHIP Mosaics containing fewer substrates can be produced upon request.
<strong>EUROIMMUN</strong> Medizinische<br />
Labordiagnostika<br />
AG<br />
BIOChIP Mosaics for Infectious Serology<br />
(For formats see pages 144/145)<br />
FI 282510011 g FI 282610011 g<br />
MYOCARDITIS PROFILE 1 INFECTIO<strong>US</strong> ARThRITIS PROFILE 1<br />
Igg IgM Igg IgM<br />
Field A 1 : 10 1 : 10 Field A 1 : 10 1 : 10<br />
V Verification BIOCHIP*** V Verification BIOCHIP***<br />
1. Mumps virus 1. VZV<br />
2. Adenovirus type 3 2. Influenza virus type A (H1N1)<br />
3. Influenza virus type A (H1N1) 3. Influenza virus type A (H3N2)<br />
4. Influenza virus type A (H3N2) 4. Influenza virus type B<br />
Field B 1 : 10 1 : 10 Field B 1 : 10 1 : 10<br />
5. Influenza virus type B 5. Yersinia enterocolitica O:3* , **<br />
6. Parainfluenza virus type 1 6. Yersinia enterocolitica O:6* , **<br />
7.. Parainfluenza virus type 2 7.. Yersinia enterocolitica O:9* , **<br />
8. Mycoplasma pneumoniae 8. Toxoplasma gondii**<br />
Field C 1 : 100 1 : 10 Field C 1 : 100 1 : 10<br />
9. CMV** 9. Borrelia afzelii<br />
10. Coxsackie virus type B1 10. Borrelia burgdorferi sensu stricto (CH)<br />
11. Coxsackie virus type A16 11. Borrelia garinii<br />
12. Echo virus type 7. 12. Chlamydia trachomatis**<br />
Field D 1 : 100 1 : 10<br />
13. Borrelia afzelii<br />
14. Borrelia burgdorferi sensu stricto (CH)<br />
15. Borrelia garinii<br />
16. Chlamydia pneumoniae<br />
RSV<br />
Adenovirus<br />
type 3<br />
Influenza<br />
virus type A<br />
(H3N2)<br />
Influenza<br />
virus type A<br />
(H1N1)<br />
Influenza<br />
virus type B<br />
Parainfluenza<br />
virus type 1<br />
Parainfluenza<br />
virus type 3<br />
Parainfluenza<br />
virus type 2<br />
Parainfluenza<br />
virus type 4<br />
Bordetella<br />
pertussis<br />
Mycoplasma<br />
pneumoniae<br />
Bordetella<br />
parapertussis<br />
* For clinical evaluation the results must be confirmed with a CE approved test.<br />
** For practical reasons the recommended incubation differs from the standard incubation for this substrate.<br />
*** Field A contains a verification BIOCHIP. The incubation was performed correctly if the dots show a visible colour reaction.<br />
BIOCHIP Mosaics containing fewer substrates can be produced upon request.<br />
Coxsackievirus<br />
type B1<br />
Coxsackievirus<br />
type A7<br />
Chlamydia<br />
pneumoniae<br />
Echovirus<br />
type 7<br />
Haemophilus<br />
influenzae<br />
Klebsiella<br />
pneumoniae<br />
Legionella<br />
pneumophila<br />
serotype 1<br />
Legionella<br />
pneumophila<br />
serotype 12<br />
— 63 —<br />
<strong>EUROIMMUN</strong> <strong>Product</strong>s for<br />
Infectious Serology
<strong>EUROIMMUN</strong> Medizinische<br />
Labordiagnostika<br />
AG<br />
<strong>EUROIMMUN</strong> <strong>Product</strong>s for<br />
Infectious Serology<br />
— 64 —<br />
absorbent<br />
Additional Reagents for the Determination of Acute Infections<br />
EUROSORB IgG/RF Absorbent.<br />
Order No. Formats<br />
ZF 127.0-0145 page 150<br />
rheumatoid factor<br />
IgG<br />
Reagents for determination of low-avidity antibodies<br />
in infectious serology.<br />
Order No. Formats<br />
ZF 1130 und ZF 1131 page 149<br />
IgG Absorpion<br />
• Before a patient’s serum is tested for specific antibodies of the IgM class, antibodies<br />
of class IgG must be removed, for example by immunoabsorption.<br />
• Specifically bound IgG would displace IgM from the antigen leading to false IgM<br />
negative test results.<br />
• Moreover, the absorption prevents any IgM class rheumatoid factors present<br />
from reacting with specifically bound IgG and thus leading to false IgM positive<br />
test results.<br />
• Indication: an IgG absorption of serum samples should always be performed<br />
for all IgM antibody determinations in infectious serology before incubating the<br />
sera.<br />
• IgG/RF absorbent is contained in the sample buffer in all ELISAs for infectious<br />
serology (class IgM).<br />
EUROSORB IgG/RF Absorbent for Indirect Immunofluorescence<br />
• Functional principle: the EUROSORB reagent contains an anti-human IgG antibody<br />
preparation from goat. Immunoglobulin G of a serum or plasma sample is<br />
bound with high specificity by these antibodies and precipitated. If the sample<br />
also contains rheumatoid factors, these will be absorbed by the anti-human<br />
IgG/IgG complex.<br />
• Incubation time of the sample with the reagent is 15 minutes.<br />
• All IgG subclasses are bound and precipitated by the anti-human IgG antibodies.<br />
• Human serum IgG in concentrations of up to 15 mg/ml and rheumatoid factors<br />
are completely removed by the absorbent (average se rum IgG concentration in<br />
adults: 12 mg/ml).<br />
• The recovery rate of the IgM fraction is almost 100%.<br />
• One unit contains 4.5 ml absorbent, sufficient for the absorption of 100 serum<br />
samples.<br />
Urea Solutions and Avidity Buffers for the Determination of Low-<br />
Avidity Antibodies in Infectious Serology<br />
• To identify low-avidity antibodies in a patient’s serum, two immunofluorescence<br />
tests are performed in parallel: one test is carried out in the conventional way,<br />
the other one includes urea treatment between incubations with patient’s serum<br />
and per oxidase-labelled anti-human IgG, resulting in the detachment of lowavidity<br />
antibodies from the antigens.<br />
• Low-avidity antibodies are present if the fluorescence intensity is significantly<br />
reduced (two intensity levels ore more) by urea treatment.<br />
• The following reagents for avidity determination are available:<br />
IFT, Ab against Order No. Avidity Solution Incubation Time<br />
Rubella ZF 1130-0501 urea solution, 5 M 10 min<br />
WNV ZF 1130-0601 urea solution, 6 M 10 min<br />
Toxopl. ZF 1130-0801 urea solution, 8 M 10 min<br />
EBV-EA, EBV-CA ZF 1130-0801 urea solution, 8 M 30 min<br />
CMV ZF 1131-0101-1 avidity buffer 1 10 min<br />
VZV ZF 1131-0101-2 avidity buffer 2 30 min
<strong>EUROIMMUN</strong> Medizinische<br />
Labordiagnostika<br />
AG<br />
<strong>EUROIMMUN</strong> PRODUCTS FOR ALLERgOLOgY<br />
— 65 —<br />
<strong>EUROIMMUN</strong> <strong>Product</strong>s for<br />
Allergology
<strong>EUROIMMUN</strong> Medizinische<br />
Labordiagnostika<br />
AG<br />
<strong>EUROIMMUN</strong> <strong>Product</strong>s for<br />
Allergology<br />
— 66 —<br />
Grasses<br />
Trees<br />
Weeds<br />
Mites<br />
Animal hair<br />
Mould spores<br />
CCD<br />
Indicator<br />
Incubated EUROLINE Allergy Profile Inhalation.<br />
Order No. Formats<br />
DP ####-1601 E Page 83<br />
Incubated ELISA Total IgE.<br />
Order No. Formats<br />
EV 3840-9601 E Page 100<br />
Allercoat TM 6 ELISA.<br />
Order No. Formats<br />
EP ####-0110 E Page 101-120<br />
ZP ####-#### E<br />
Antibodies of Class IgE against Allergens<br />
EUROLINE: Specific IgE<br />
• Determination of specific IgE in serum.<br />
• Indication: Clarification of allergic reactions to inhalation allergens, food<br />
allergens and insect venom allergens.<br />
• Serum dilution: undiluted (or 1 : 10); conjugate class anti-IgE (monoclonal), APlabelled.<br />
• Antibodies against up to 36 allergens per strip can be simultaneously monospecifically<br />
detected.<br />
• EUROLINE profiles with different allergen compositions are available for various<br />
test requirements: atopy, inhalation, food, insect venoms and cross reactions.<br />
• Molecular allergy diagnostics based on single purified allergen components<br />
(SPAC) allows precise identification of the individual cause of an allergic<br />
reaction.<br />
• The incubated EUROLINE test strips are scanned using a flatbed scanner. The<br />
EUROLine Scan programme recognises the position of the strips, identi fies the<br />
bands and measures their inten sity.<br />
<strong>EUROIMMUN</strong> Microplate ELISA: Total IgE<br />
• Determination of the total IgE concentration in serum.<br />
• Indications: Differentiation between allergic and intrinsic asthma, between<br />
allergic and vasomotor rhinitis, and between atopic and seborrhoic dermatitis.<br />
• Serum dilution 1 : 10; conjugate class anti-IgE (monoclonal), POD-labelled.<br />
• One microplate well incubated per patient.<br />
• 4-point calibration, quantitative.<br />
• Coating: anti-human IgE, polyclonal.<br />
• The Total IgE ELISA serves as a screening test for allergy diagnostics and<br />
provides an indication for the presence of an allergic reaction.<br />
Allercoat 6 Microplate ELISA: Specific IgE<br />
• Determination of allergen-specific IgE concentrations in serum.<br />
• Indication: identification of allergic reactions to more than 700 different allergens<br />
and allergen mixtures.<br />
• Serum dilution: undiluted, conjugate class anti-human IgE, AP-labelled.<br />
• One microplate well per allergen/allergen mixture is incubated for each patient.<br />
• Calibration: 6-point calibration, quantitative; using reference preparation 2 IRP<br />
7.5/502 from the WHO.<br />
• The allergens are coupled to paper rings and can be flexibly configured for each<br />
sample according to the analysis.<br />
• Customized pre-assembled microplates with individual allergy parameters are<br />
available on request (Order no.: EP 9901-0101). Orders can be made online with<br />
the provided software. Please contact us!<br />
• A light table and special evaluation software are available to supplement the<br />
simple manual Allercoat 6 ELISA procedure.<br />
• Allercoat 6 ELISA are automatable using the <strong>EUROIMMUN</strong> Analyzer I and the<br />
<strong>EUROIMMUN</strong> Allercoat software.
<strong>EUROIMMUN</strong> Medizinische<br />
Labordiagnostika<br />
AG<br />
Antibodies of Class IgE against Allergens<br />
Customized Laboratory Software for Flexible Automation of<br />
Allercoat System<br />
• Convenient online ordering of individual pre-prepared Allercoat microtiter<br />
plates.<br />
• Integrated light table for manual preparation of ELISA plates and sample<br />
distribution.<br />
• Direct control of photometer, automated evaluation via calibration curve and<br />
compilation of results.<br />
• Fully automated incubation of samples and evaluation of results via connection<br />
to the <strong>EUROIMMUN</strong> Analyzer I.<br />
• Fully automated administration, documentation and archiving of all data.<br />
• Simple operation using graphic user commands and clear presentation of the<br />
most important information at a glance.<br />
• Connection to commonly used laboratory systems for convenient transfer of<br />
requests and results.<br />
• Additional security through user administration with individual access rights<br />
and confirmation step before results editing.<br />
Individual equipment<br />
with allergen rings<br />
Pipette: 50 µl per well<br />
Incubate: 60 min, 37. °C<br />
Wash: 300 µl wash buffer<br />
per well<br />
Pipette: 50 µl per well<br />
Incubate: 60 min, 37. °C<br />
Wash: 300 µl wash buffer<br />
per well<br />
Pipette: 100 µl per well<br />
Incubate: 30 min, 37. °C<br />
Pipette: 100 µl per well<br />
Evaluate: photometric measurement<br />
at a wavelength of 405 nm<br />
allergen rings<br />
undiluted<br />
samples<br />
enzyme<br />
conjugate<br />
chromogen-<br />
/substrate sol.<br />
stop<br />
solution<br />
Allercoat TM Software.<br />
Fitting of allergen rings into<br />
microplate wells<br />
— 67. —<br />
<strong>EUROIMMUN</strong> <strong>Product</strong>s for<br />
Allergology
<strong>EUROIMMUN</strong> Medizinische<br />
Labordiagnostika<br />
AG<br />
general Delivery Conditions<br />
<strong>Product</strong>s from <strong>EUROIMMUN</strong> will be delivered according to the following conditions, as long as no other conditions have been agreed in<br />
writing. On issuing an order, the buyer acknowledges the delivery conditions. <strong>EUROIMMUN</strong> does not accept the sales conditions of the<br />
buyer, even when <strong>EUROIMMUN</strong> has not expressly contradicted them. Contract terms are valid for all future business connection even if<br />
they have not been expressly agreed upon again.<br />
Orders: Orders to <strong>EUROIMMUN</strong> can be made in writing (including electronic communication) or verbally. Verbally issued orders first<br />
become legally binding for <strong>EUROIMMUN</strong> with a written confirmation from the orderer or with the delivery of goods.<br />
Delivery: <strong>EUROIMMUN</strong> Medizinische Labordiagnosika AG (<strong>EUROIMMUN</strong>) generally delivers to end users within 7. days of order receipt and<br />
to distributors within 14 days, but is not tied to any definite delivery period. If a delivery is not possible through unforseeable circumstances<br />
(e.g., factory disruptions, raw material delivery delays, transport difficulties, strikes), the delivery obligation does not apply. EURO IMMUN<br />
reserves the right to deliver in part. The delivery obligation of <strong>EUROIMMUN</strong> ceases as long as the customer is in arrears. <strong>EUROIMMUN</strong><br />
chooses the packing and dispatching method at its own discretion, according to the respective requirements.<br />
<strong>Product</strong> characteristics: The delivered products comply with specifications given in the product catalogue, on the product itself, or in<br />
the supplied information sheets. If specifications are inconsistent, the labels on the product itself and the details in the information sheet<br />
provided with the product apply. After the given expiration date has passed, the test reagents must not be used.<br />
<strong>EUROIMMUN</strong> products must only be used for the intended purpose. It is the buyer’s responsibility to check the functional capability<br />
immediately on receipt of the product as well as on every day of usage. In particular, the functioning of test reagents can be perturbed<br />
through causes outside the direct influence of the maunfacturer, for example, through suboptimal transportation, incorrect storage,<br />
or incorrect usage. When test reagents delivered from <strong>EUROIMMUN</strong> are used, the user must supervise the analysis with appropriate<br />
proficiency.<br />
Warranty and liability: If nothing else has been agreed upon in writing, all risks pass to the buyer as soon as the goods have been<br />
delivered to the carrier or has left <strong>EUROIMMUN</strong> premises for dispatch. With notification of dispatch by <strong>EUROIMMUN</strong> the risk passes to the<br />
buyer if the delivery is postponed or delayed by request of the buyer. On receipt of the goods, the buyer has one working day to check if<br />
they are in accordance with the nature and quantity of the agreement and if the goods are free from visible defects. If any complaints arise<br />
from this inspection, they should be communicated to <strong>EUROIMMUN</strong> in writing within 2 working days. Hidden defects or functional faults<br />
that were not identifiable with the initial inspection and are later discovered should be communicated to <strong>EUROIMMUN</strong> in writing within<br />
14 days of receipt of the goods. If no complaints are received within the stipulated period, it is assumed that the goods are of appropriate<br />
quality and quantity for the customer. Complaints do not release the buyer from payment obligation.<br />
With prompt and reasonable complaints on the grounds of product defects or the delivery of something other than the ordered goods,<br />
<strong>EUROIMMUN</strong> is obliged to exchange or amend the goods within 14 days or to withdraw or reimbourse the payment, as it chooses. If the<br />
defect is not corrected in spite of delivery of a replacement or an amended product, the buyer can demand that the sale is cancelled.<br />
If defects are promptly reclaimed, <strong>EUROIMMUN</strong> has the choice between a further delivery or an appropriate credit note. Further compensatory<br />
claims from the buyer are excluded, as long as <strong>EUROIMMUN</strong> has not violated its contractual or legal obligations through outright negligence<br />
or by intention. In such cases <strong>EUROIMMUN</strong> provides compensation of up to a maximum of 20 times the price of one packet (test kit) of<br />
the defective product. <strong>EUROIMMUN</strong> takes no responsibility for any damage resulting from faulty goods.<br />
Prices: Prices from the official <strong>EUROIMMUN</strong> pricelist in the country of the buyer are applicable. Invoices are provided in the agreed<br />
currency. Prices are “ex works”. Expenses for packing and freight as well as for cooling during transport are added on, including costs for<br />
the disposal of packaging material by the customer. Statutory value added tax is not included in the list prices.<br />
Payment terms: Payment obligations in countries of export must be settled within 30 days after recieving the goods. No cash discounts<br />
will be given unless agreed in writing. Payments by cash transfer or check are valid from the timepoint that the invoice amount is credited<br />
to a <strong>EUROIMMUN</strong> bank account. In cases of payment arrears, <strong>EUROIMMUN</strong> reserves the right to charge compensatory interest of 10% p.<br />
a., applicable from the settlement date, without any further notice. In special cases <strong>EUROIMMUN</strong> can arrange alternative payment periods<br />
or demand advance payments. <strong>EUROIMMUN</strong>’s demands resulting from an order cannot be offset by the customer through counter<br />
demands.<br />
Ownership rights: The delivered goods remain the property of <strong>EUROIMMUN</strong> until full payment. Selling on of <strong>EUROIMMUN</strong> products is<br />
only permitted for companys who are authorized in writing from <strong>EUROIMMUN</strong> to do so. Software received from <strong>EUROIMMUN</strong> should not<br />
be passed on to third parties without written permission from <strong>EUROIMMUN</strong>.<br />
Consultation: Advice from <strong>EUROIMMUN</strong>, although provided to the best knowledge, is nevertheless not binding. No liability claims can<br />
ensue from erroneous advice.<br />
Applicable law, place of jurisdiction, ineffective regulations: The contract conditions are subject to the laws of the Federal Republic<br />
of Germany. The United Nations Conventions on Contracts for the International Sale of Goods does not apply. The place of jurisdiction<br />
and fulfilment is Luebeck. If any provision of these general delivery conditions will be held invalid or unenforcable, this general delivery<br />
condition will not be rendered invalid as a whole, and the provisions will be changed and interpreted so as best to accomplish the objective<br />
of the unenforcable or invalid provision.<br />
— 151 —
<strong>EUROIMMUN</strong> Medizinische<br />
Labordiagnostika<br />
AG<br />
— 152 —<br />
Autoantibody Diagnostics<br />
acetylcholine receptor (ACHR) ...................................................................................................................12, 17., 93<br />
actin ................................................................................................................... 16, 17., 34, 40, 50, 7.2, 126, 132-133<br />
adrenal cortex ............................................................................................................................... 13, 17., 47., 7.0, 121<br />
AGNA ...................................................................................................................................................................... 17.<br />
alpha-amylase ...............................................................................................................................................100, 117.<br />
AMA ......................................................... 13, 16-17., 26, 35, 39-42, 47., 50, 7.2, 80, 82, 91, 121, 126, 130, 132-133<br />
AMPA receptor ....................................................................................................................................12, 17., 45, 123<br />
amphiphysin ........................................................................................................................................17., 44, 82, 122<br />
ANA ...............................................................14, 16-17., 26, 34-36, 39-40, 44, 48, 50, 7.2, 82, 90, 124-126, 130-133<br />
ANCA ..................................................................................................12-14, 16-17., 34, 48-50, 7.0, 89, 123-125, 127.<br />
aquaporin-4 ............................................................................................................................12, 17., 44, 7.0, 122-123<br />
ASMA ................................................................................................................... 12, 16-17., 39-40, 7.2, 126, 132-133<br />
basement membrane .................................................................................. 12, 14, 16-17., 43, 7.1, 82, 89, 125, 128<br />
ß2-glycoprotein ................................................................................................................................................ 16, 91<br />
bile ducts ...........................................................................................................................................................17., 34<br />
BP180 ...................................................................................................................................... 12, 17., 43, 7.1, 90, 128<br />
BP230 ...................................................................................................................................... 12, 17., 43, 7.1, 89, 128<br />
BPI ..........................................................................................................................................................17., 48, 50, 89<br />
brain ......................................................................................................................................................... 12, 121-122<br />
C1q .................................................................................................................................................................... 16, 91<br />
cANCA.....................................................................................................12, 16-17., 34, 48, 50, 7.0, 89, 123-125, 127.<br />
cardiolipin (AMA M1) ............................................................................................................................16-17., 86, 91<br />
cartilage ............................................................................................................................................................ 13, 16<br />
CASPR2 ..................................................................................................................................................14, 17., 45, 7.1<br />
cathepsin G ...........................................................................................................................................17., 48, 50, 89<br />
cell nuclei (ANA global test) .....................................................................................................................16-17., 130<br />
centromere .........................................................................................................................14, 16, 34, 36, 50, 7.2, 90<br />
cerebellum .............................................................................................................................. 12-14, 44-46, 121-122<br />
circulating immune complexes (CIC) ............................................................................................................. 16, 91<br />
collagen VII ...............................................................................................................................13, 16-17., 43, 7.3, 136<br />
CUZD1 (rPAg1) ..............................................................................................................................12, 17., 49, 7.1, 127.<br />
CV2 .......................................................................................................................................................17., 44, 82, 122<br />
cyclic citrullinated peptide (CCP) .............................................................................................................. 16, 38, 90<br />
cyclin I (PCNA) ....................................................................................................... 12, 16, 26, 34-35, 50, 7.2, 82, 90<br />
cyclin II (mitosin) ....................................................................................................................................... 12, 50, 7.2<br />
cytoplasm of granulocytes ..............................................................12-14, 16-17., 34, 48-50, 7.0, 89, 123-125, 127.<br />
desmoglein ............................................................................................................................. 12, 17., 43, 7.1, 89, 128<br />
desmosomes .................................................................................................................................12, 17., 43, 7.1, 128<br />
dsDNA ................................................................. 12, 16-17., 26, 34-37., 40, 44, 50, 7.2, 80, 82, 90, 93, 132, 148-149<br />
elastase ..................................................................................................................................................17., 48, 50, 89<br />
elastin .....................................................................................................................................................12, 16-17., 7.3<br />
ENA ....................................................................................................................................16, 24, 34, 36, 80, 82, 90<br />
endomysium...................................................................................................... 12, 17., 34, 52, 7.3, 91, 128, 134-135<br />
endothelial cells .............................................................................................................................12, 16-17., 34, 136<br />
envoplakin ...........................................................................................................................................17., 43, 89, 128<br />
epidermis .............................................................................................................................................12, 17., 7.1, 128<br />
EUROArray ...........................................................................................................................................30, 31, 68, 92<br />
EUROPL<strong>US</strong> .....................................7., 12, 34, 39-40, 42-43, 48, 50, 52, 121, 124-125, 127., 130-132, 134-135, 147.<br />
eye antigens ................................................................................................................................................... 12, 123<br />
F-actin....................................................................................................................... 16, 17., 40, 50, 7.2, 126, 132-133<br />
FT3 ...............................................................................................................................................................17., 42, 93<br />
FT4 ...............................................................................................................................................................17., 42, 93<br />
GABAB receptor ............................................................................................................................12, 17., 45, 7.0, 123<br />
GAD ............................................................................................................................. 12, 17., 44, 46, 7.0, 89, 93, 121<br />
gangliosides ......................................................................................................................................................17., 82<br />
gliadin ...........................................................................................7., 12, 17., 52, 7.3, 7.9, 100, 106, 128, 134-135, 146<br />
glomerular basement membrane (GBM).......................................................... 12, 17., 48, 50, 7.1, 82, 89, 125-126<br />
glutamate receptor ................................................................................................................12, 17., 45, 7.0, 122-123<br />
glutamic acid decarboxylase .................................................................................... 12, 17., 44, 46, 7.0, 89, 93, 121<br />
glycine receptor ..................................................................................................................................................... 17.<br />
Golgi apparatus .............................................................................................................................12, 16, 34, 50, 7.2<br />
goblet cells ....................................................................................................................................12, 17., 49, 7.1, 127.<br />
GP2 (rPAg2) ....................................................................................................................................12,17., 49, 7.1, 127.<br />
granulocyte cytoplasm ......................................................................12-14, 16-17., 34, 48-50, 7.0, 89, 123-125, 127.<br />
heart muscle ....................................................................................................................................... 13, 17., 127.-128<br />
HEp-2-cells .................................6, 9, 12-14, 34, 39-40, 44, 48, 50, 87., 120, 122, 124-125, 126, 128, 130-133, 147.<br />
histones .................................................................................................................. 13, 16, 26, 34-36, 50, 80, 82, 90<br />
Hu ............................................................................................................................................ 13, 17., 44, 7.0, 82, 122<br />
HUVEC ............................................................................................................................................................ 12, 136<br />
hypothalamus antigens ................................................................................................................................ 13, 122<br />
IA-2 .........................................................................................................................................................17., 46, 89, 93<br />
insulin .................................................................................................................................................. 17., 46, 93, 101<br />
intestinal goblet cells ...................................................................................................................12, 17., 49, 7.1, 127.<br />
intrinsic factor .................................................................................................................. 13, 17., 47., 7.1, 89, 126-127.<br />
IRMA .......................................................................................................................................................... 32, 42, 93<br />
Jo-1 ............................................................................................... 13, 16, 24, 26, 34-36, 50, 7.2, 80, 82, 90-91, 131<br />
keratin .......................................................................................................................... 12, 13, 16-17., 38, 43, 7.1, 129<br />
Ku ........................................................................................................................................13, 16, 26, 35, 50, 7.2, 82<br />
lactoferrin .................................................................................................................. 12, 17., 48-50, 7.0, 89, 125, 127.<br />
LC-1 ................................................................................................................................17., 26, 34, 40-41, 80, 82, 89<br />
Leydig cells ................................................................................................................................... 17., 47., 7.0, 121-122<br />
LGI1 ................................................................................................................................................14, 17., 45, 7.1, 128<br />
liver-kidney microsomes (LKM) ....................................... 13, 17., 26, 34, 39-41, 7.1, 80, 82, 89, 121, 126, 130-133<br />
liver-specific antigens 12-13, 17., 40-41, 7.1, 80, 82, 89, 126<br />
LKM-1 .................................................................................................................................. 17., 26, 34, 41, 80, 82, 89<br />
lung alveoli ......................................................................................................................................................17., 126<br />
lymphocyte antigens ................................................................................................................................13, 17., 125<br />
M2 antigen (AMA-M2) .................................................................................13, 17., 26, 34-35, 39-41, 7.2, 80, 82, 91<br />
M4 antigen ......................................................................................................................................13, 17., 34, 39, 80<br />
M9 antigen ......................................................................................................................................13, 17., 34, 39, 80<br />
Ma1/Ma2 ................................................................................................................................................................. 17.<br />
Mab ......................................................................................................................................................17., 42, 7.0, 121<br />
MAG .......................................................................................................................................................13, 17., 44, 7.0<br />
Mi-2 ...........................................................................................................................................13, 16, 26, 35, 50, 82<br />
Microarray ................................................................................................................................................... 30-31, 92<br />
mitochondria (AMA) ............................................................... 13, 16-17., 39-42, 47., 50, 7.2, 121, 126, 130, 132-133<br />
mitosin (cyclin II) ....................................................................................................................................... 12, 50, 7.2<br />
myelin ...............................................................................................................................................12-13, 17., 44, 7.0<br />
myeloperoxidase (MPO) ........................................................................... 7., 13, 17., 48, 50, 7.0, 80, 82, 89, 124-125<br />
nDNA..........................................................................12, 16-17., 26, 34-37., 40, 44, 50, 7.2, 80, 82, 90, 132, 148-149<br />
nerves ............................................................................................................................................ 12-14, 17., 44, 122<br />
neuronal autoantibodies ......................................................................................................................17., 44, 45, 82<br />
NMDA receptor ......................................................................................................................12, 17., 45, 7.0, 122-123<br />
NMO ...........................................................................................................................................................17., 44, 122<br />
nRNP/Sm ........................................................................................................ 16, 24, 26, 34-36, 50, 80, 82, 90, 131<br />
nuclear membrane................................................................................................................................17., 34, 50, 7.2<br />
nucleosomes .................................................................................................................16, 34-35, 37., 50, 80, 82, 90<br />
oesophagus ............................................................................................................ 12-13, 43, 52, 128-129, 134-135<br />
ovarial antigens ........................................................................................................................................47., 89, 121<br />
P/Q calcium channel (VGCC) ................................................................................................................................. 17.<br />
pANCA ....................................................................................................13, 16-17., 34, 48, 50, 7.0, 89, 123-125, 127.<br />
pancreas islets ........................................................................................................................14, 46-47., 7.0, 121-122<br />
parathyroid gland .................................................................................................................................... 13, 17., 121<br />
parietal cells (PCA) ..................................................................................................12, 17., 47., 7.1, 89, 121, 126-127.<br />
parotid gland excretory ducts and acini .................................................................................................14, 17., 127.<br />
PCA-2 ...................................................................................................................................................................... 17.<br />
PCNA (cyclin I) ....................................................................................................... 12, 16, 26, 34-35, 50, 7.2, 82, 90<br />
phosphatidylserine .......................................................................................................................................... 16, 91<br />
phospholipase-A2 receptor (PLA2R) ...........................................................................................17., 51, 7.1, 89, 125<br />
phospholipids ......................................................................................................................................... 16-17., 87., 91<br />
pituitary gland antigens ...........................................................................................................................13, 17., 122<br />
PL-12 ................................................................................................................................................16, 26, 35, 50, 82<br />
PL-7. ..................................................................................................................................................16, 26, 35, 50, 82<br />
placenta .....................................................................................................................................................14, 17., 121<br />
PM-Scl ........................................................................................................................... 16, 26, 34-36, 50, 80, 82, 90<br />
PNMA2 (Ma2/Ta) .........................................................................................................................................17., 44, 82<br />
proteinase 3 (PR3) ......................................................................................7., 14, 17., 48, 50,7.0, 80, 82, 89, 124-125<br />
renal glomeruli (GBM)........................................................................................ 12, 17., 48, 50, 7.1, 82, 89, 125-126<br />
recoverin ......................................................................................................................................................17., 44, 82<br />
reticulin .......................................................................................................................................14, 17., 128, 134-135<br />
retina .....................................................................................................................................................13-14, 17., 123<br />
Index<br />
rheumatoid factors .................................................................................................................................... 16, 64, 91<br />
Ri ....................................................................................................................................... 14, 17., 38, 44, 7.0, 82, 122<br />
RIA ............................................................................................................................................. 16, 32, 37., 42, 46, 93<br />
ribosomal P-proteins......................................................................................................................14, 16, 34, 7.2, 91<br />
RNA-Polymerase ................................................................................................................14, 16, 31, 34, 35, 50, 82<br />
Ro-52 ............................................................................................................. 16-17., 26, 34-36, 41, 50, 80, 82, 87., 90<br />
rPAg1/2 ................................................................................................................................................ 12, 49, 7.1, 127.<br />
Sa ................................................................................................................................................................ 16, 38, 90<br />
Saccharomyces cerevisiae ............................................................................ 15, 17., 49, 7.9, 100, 105-106, 127., 146<br />
saliva ......................................................................................................................................................................100<br />
Scl-7.0 .................................................................................................14, 16, 24, 26, 34-36, 50, 7.2, 80, 82, 90, 131<br />
skeletal muscle........................................................................................................................12-14, 17., 40, 126-127.<br />
SLA/LP............................................................................................................................17., 26, 34, 40-41, 80, 82, 89<br />
Sm .......................................................................................................14, 16, 24, 26, 34-36, 50, 7.2, 80, 82, 90, 131<br />
smooth muscles (ASMA) ................................................................................... 12, 16-17., 39-40, 7.2, 126, 132-133<br />
SOX1 ................................................................................................................................................................. 44, 82<br />
Sp100 ........................................................................................................................................ 17., 26, 40-41, 7.2, 82<br />
spermatozoa ...........................................................................................................................14, 17., 7.0, 89, 121-122<br />
spindle fibers........................................................................................................................................16, 34, 50, 7.2<br />
SS-A ...........................................................................14, 16-17., 24, 26, 34-36, 50, 7.2, 80, 82, 87., 90, 130-131, 147.<br />
SS-B ............................................................................... 14, 16-17., 24, 26, 34-36, 50, 7.2, 80, 82, 90, 130-131, 147.<br />
ssDNA .........................................................................................................................................14, 16-17., 36, 50, 90<br />
striated muscles ......................................................................................................................................... 14, 40, 7.1<br />
TAb .......................................................................................................................................................17., 42, 7.0, 121<br />
testis .................................................................................................................................. 14, 17., 42, 47., 7.0, 121-122<br />
thrombocyte antigens ..............................................................................................................................14, 17., 125<br />
thyroglobulin (TG) ................................................................................................14, 17., 42, 47., 7.0, 80, 89, 93, 121<br />
thyroid gland ...................................................................................................13, 14, 17., 42, 47., 7.0, 80, 89, 93, 121<br />
thyroid peroxidase (TPO) ...............................................................................................................17., 42, 80, 89, 93<br />
titin ...............................................................................................................................................................17., 44, 82<br />
Tr ............................................................................................................................................................................. 17.<br />
TRAb ......................................................................................................................................................17., 42, 89, 93<br />
transglutaminase .............................................................................................. 12, 17., 34, 52, 7.3, 91, 128, 134-135<br />
U1-nRNP ............................................................................................................................................... 14, 16, 7.2, 90<br />
vascular endothelium .................................................................................................................................16, 17., 7.3<br />
VGKC ....................................................................................................................................................14, 17., 45, 128<br />
vimentin ................................................................................................................................................ 14, 16, 50, 7.2<br />
Yo ......................................................................................................................................14, 17., 44, 7.0, 82, 121-122<br />
zink transporter 8 ................................................................................................................................................... 17.<br />
zona pellucida .................................................................................................................................................. 7.0, 89<br />
hormones and Proteins<br />
alpha-amylase ...............................................................................................................................................100, 117.<br />
cortisol ...................................................................................................................................................................100<br />
vitamin D ...............................................................................................................................................................100<br />
Infectious serology<br />
Adenovirus ...............................................................................................................................15, 62-63, 7.7., 98, 142<br />
Bartonella ....................................................................................................................................15, 18, 62, 7.5, 139<br />
Bordetella .........................................................................................................................15, 18, 62, 7.3, 83, 94, 137.<br />
Borrelia ...................................................................................... 15-18, 21, 28, 54-55, 62-63, 7.4, 83, 87., 94-95, 137.<br />
Brucella abortus ..................................................................................................................................................... 95<br />
Campylobacter .............................................................................................................................15, 18, 7.3, 94, 137.<br />
Candida ................................................................................................................................. 15, 18, 62, 7.9, 113, 145<br />
Chikungunya virus ..................................................................................................................... 15, 61, 7.9, 100, 145<br />
Chlamydia ............................................................................................ 15-16, 18, 60, 62-63, 7.4-7.5, 88, 95, 138-139<br />
Coxsackie virus ..........................................................................................................15, 18, 62-63, 7.8, 99, 142-143<br />
Crimean Congo fever virus ............................................................................................................... 15, 61, 7.9, 144<br />
Cytomegalovirus (CMV) .......................................................................7., 15, 21, 62-64, 7.6, 83, 88, 96-97., 140, 149<br />
Dengue virus .................................................................................................................................15, 61, 7.7., 98, 142<br />
EBNA .......................................................................................................................... 15, 18, 56-57., 7.9, 83, 100, 144<br />
EBV-CA .................................................................................... 7., 15, 18, 21, 56-57., 62, 64, 68, 7.8, 99, 100, 143-144<br />
EBV-EA ........................................................................................................................ 7., 18, 56, 62, 64, 7.9, 100, 144<br />
Echinococcus granulosus ...........................................................................................................15, 7.5, 88, 115, 140<br />
Echo virus .........................................................................................................................15, 18, 62-63, 7.8, 142-143<br />
Epstein-Barr virus (EBV) ..............................7., 15, 18, 21, 56-57., 62, 64, 68, 7.8-7.9, 83, 88, 99, 100, 143-144, 150<br />
EUROPL<strong>US</strong> .................................................................................................................................... 7., 54, 56, 137., 144<br />
EUROSORB IgG/RF absorbens ..................................................................................................................... 68, 150<br />
Haemophilus influenzae ........................................................................................................................... 15, 18, 62<br />
Hantavirus ..............................................................................................................................15, 61, 7.8, 83, 99, 143<br />
Helicobacter pylori ........................................................................................................... 15, 18, 58, 7.3, 87., 94, 137.<br />
Herpes simplex (HSV) ............................................................................. 15, 18, 21, 59, 62, 7.5-7.6, 83, 88, 96, 140<br />
HEV ......................................................................................................................................................................... 96<br />
HHV-6 ............................................................................................................................................ 15, 18, 62, 7.6, 140<br />
HIV ..................................................................................................................................................................... 18, 62<br />
Influenza virus ....................................................................................................................15, 62-63, 7.7.-7.8, 99, 142<br />
Japanese encephalitis virus .........................................................................................................15, 61, 7.7., 98, 141<br />
Klebsiella pneumoniae .............................................................................................................................. 15, 18, 62<br />
Legionella ...............................................................................................................................15, 18, 62, 7.4, 95, 138<br />
Leishmania ...................................................................................................................................15, 18, 61, 7.5, 140<br />
Listeria ........................................................................................................................................ 15, 18, 62, 7.4, 137.<br />
Measles virus ....................................................................................................................15, 18, 21, 62, 7.6, 97., 141<br />
Mumps virus ............................................................................................................... 15, 18, 21, 62-63, 7.6, 97., 141<br />
Mycoplasma .................................................................................................................15, 18, 62-63, 7.5, 95-96, 139<br />
Parainfluenza virus .................................................................................................................15, 62-63, 7.8, 99, 142<br />
Parvovirus ......................................................................................................................................................... 83, 97.<br />
Plasmodium ......................................................................................................................................7., 15, 18, 61, 7.5<br />
Respiratory syncytial virus (RSV) ..........................................................................................15, 18, 62, 7.7., 98, 142<br />
Rift valley fever virus.................................................................................................................. 15, 61, 7.9, 143-144<br />
Rubella virus ........................................................................................... 15, 18, 21, 62, 64, 7.6, 83, 88, 97., 140-141<br />
Sandfly fever virus............................................................................................................................. 15, 61, 7.8, 143<br />
SARS coronavirus .................................................................................................................................... 15, 61, 141<br />
TBE virus ....................................................................................................................15, 18, 21, 54, 61, 7.6, 98, 141<br />
Toxoplasma gondii ...........................................................................................7., 15, 18, 21, 62-63, 7.5, 83, 96, 140<br />
Treponema pallidum .................................................................................................. 15, 18, 62, 7.3-7.4, 87., 94, 137.<br />
Ureaplasma urealyticum ............................................................................................................................... 18, 139<br />
Usutu virus ....................................................................................................................................................... 61, 98<br />
Varicella zoster virus (VZV) .................................................................................7., 15, 21, 62-64, 7.6-7.7., 97.-98, 141<br />
VlsE ................................................................................................................7., 15, 28, 54-55, 7.4, 83, 87., 94-95, 137.<br />
West Nile virus ..............................................................................................................7., 15, 21, 61, 64, 7.7., 98, 141<br />
Yellow fever virus ...............................................................................................................................15, 61, 7.7., 141<br />
Yersinia enterocolitica ..................................................................................................... 15-16, 18, 63, 88, 95, 138<br />
Allergology<br />
animal allergens ......................................................................................................................................19, 103-104<br />
environmental allergens ..........................................................................................................................19, 117.-118<br />
food ....................................................................................................................................19, 66, 81, 84-85, 105-111<br />
further allergens....................................................................................................................................................115<br />
grasses .......................................................................................................................................................19, 66, 112<br />
herbal and flower pollen .....................................................................................................19, 66, 115-116, 119-120<br />
house dust .......................................................................................................................................................19, 113<br />
insects ............................................................................................................19, 41, 52, 66, 81, 86, 89-91, 100, 113<br />
mites ..................................................................................................................................................19, 66, 103, 113<br />
moulds ...................................................................................................................................................... 19, 113-115<br />
parasites ....................................................................................................................................................18, 19, 115<br />
pharmaceutical drugs .............................................................................................................................. 19, 101-102<br />
trees .......................................................................................................................................................... 19, 115-116<br />
Automation<br />
EUROBlotMaster ....................................................................................................... 26-27., 29, 35, 41, 44-45, 57.-59<br />
EUROBlotOne ......................................................................................................................................................... 27.<br />
<strong>EUROIMMUN</strong> Analyzer ............................................................................................................................... 22, 66-67.<br />
EUROLineScan ................................................................................................ 26-29, 35, 41, 44, 55, 57.-59, 66, 149<br />
EUROPattern ...............................................................................................................9, 34, 68-69, 7.2, 130-131, 147.<br />
EUROStar.................................................................................................................................................................. 9<br />
IF Sprinter ................................................................................................................................................................. 8<br />
instruments .................................................................................................................................................8, 22, 27.<br />
Sprinter XL ............................................................................................................................................................... 8
One laboratory management software<br />
for all automated systems<br />
<strong>EUROIMMUN</strong> Medizinische<br />
Labordiagnostika<br />
AG<br />
Pos.<br />
Neg.<br />
Borderline<br />
?<br />
Pos.<br />
Neg.<br />
Borderline<br />
?<br />
Repeat<br />
Pos.<br />
Neg.<br />
Borderline<br />
?<br />
Further<br />
diagnostics<br />
Macros<br />
YG_0250_R_UK_B05, 9/2012<br />
Computer result<br />
+++ Centromeres 1:3,200<br />
Visual microscopic result<br />
++++ Centromeres 1:10,000<br />
Addition:<br />
ENA EUROPL<strong>US</strong> neg.<br />
Final result<br />
+++ Centromeres 1:3,200<br />
Unofficial remark<br />
98% Homogeneous pattern<br />
Granular pattern<br />
Nucleolar pattern<br />
Centromeres<br />
1:3,200 94%<br />
Nuclear dots<br />
Mitosis pattern<br />
Cytoplasmic pattern<br />
Negative<br />
Nuclear membrane<br />
Ku<br />
PCNA<br />
Delete all X Reset Mitosin (CENP F)<br />
Coilin (Few nuclear dots)<br />
Spindle apparatus<br />
Centrioles<br />
Jo-1<br />
Other<br />
IIFT<br />
Verify synthesis<br />
15/28<br />
Serum No.: 127<br />
ELISA<br />
Immunoblots<br />
?<br />
EUROLabOffice<br />
<strong>EUROIMMUN</strong> AG · D-23560 Luebeck (Germany) · Seekamp 31 · Tel +49 451 58550 · Fax 5855591 · E-mail euroimmun@euroimmun.de<br />
D-23560 Luebeck (Germany) · Seekamp 31 · Telephone: +49 451 58550 · Fax: +49 451 5855591 · euroimmun@euroimmun.de