ALLERGEN Total IgE REF - Radim S.p.A.

ENZYME IMMUNOASSAY FOR QUANTITATIVE DETERMINATION OF TOTAL IgE IN HUMAN
SERUM AND PLASMA
FOR IN VITRO DIAGNOSTIC USE ONLY
1.0 INTENDED USE
The ALLERGEN Total IgE is an enzyme immunoassay kit for the quantitative determination of total
Immunoglobulins of type E (IgE) in human serum or plasma. It is strictly for in vitro diagnostic use only
and the obtained results should be used in conjunction with other data available to the physician.
2.0 SUMMARY AND EXPLANATION OF THE TEST
The Immunoglobulins belonging to E class (IgE) contain 2 types of polypeptide chains, have a molecular
weight of approximatively 200.000 (1) and bind to surface of mast cells and basophilic granulocytes. The
binding of allergens to cell-bound IgE causes these cells to release histamine and other vasoactive
substances, thereby initiating the events recognized as allergic reactions (1,2,3). Most patients with
atopic allergic diseases, such as atopic asthma, atopic dermatitis and hay fever have been shown to
exhibit an increase of total IgE levels in blood (4,5,6). Non-allergic individuals have IgE concentrations
that vary widely, and during childhood they normally show increase, reaching adult levels in the second
decade of life (7,8). Measurement of total IgE concentration may be of value in the early detection of
allergy in infants, and as a means for predicting future atopic manifestations (9,10,11). Significant
elevation may be found not only in allergic patients, but also in cases of IgE myeloma, pulmonary
aspergillosis, and during the active stage of parasitic infestations (12,13). Increased levels of IgE are
found in cases of hypergammaglobulinemia, autoimmune diseases, ulcerative colitis, hepatitis, cancer
and malaria (14).
3.0 PRINCIPLE OF THE TEST
The ALLERGEN total IgE is based on the enzyme immunoassay, sandwich principle. The testing sample
is added to the monoclonal anti-human-IgE antibodies coated onto the wells of the microstrips. During
the first incubation the solid phase anti-IgE captures the sample IgE. The addition and subsequent
incubation with an Anti-IgE-Biotin conjugate lead to the formation of the sandwich: solid phase-Anti-IgE :
IgE : Anti-IgE-Biotin. The further incubations with the Streptavidin-Peroxidase conjugate and then with
the substrate chromogen, respectively, lead to a blue color development. The enzymatic reaction is then
blocked with a stop solution which turns yellow the blocked solution. The resulting colour is read
photometrically at 450 nm. The concentration of sample IgE is directly related to the colour intensity.
4.0 KIT COMPONENTS
MT PLATE
CONJ│BIOT
CAL│A…F
BUF│INC
DIL│SPE
CONJ│HRP
Reagents
Microplate sensitized with anti-IgE: one bag containing 12 strips x 8 wells,
coated with monoclonal (mouse) anti-human IgE. Store the unused strips at
2…8°C in the plastic zip pouch with the desiccant.
Anti-IgE-Biotin Conjugate: one vial (13 ml) of anti-IgE-Biotin conjugate in TRIS
buffer, pH 8.1, preservatives and stabilizers. Ready to use.
Calibrators for Total IgE: 6 Vials, 0.5 ml each. The vials contain human IgE in
horse serum with 0.01% of Proclin 300 as preservative. The IgE concentrations
are: 0 - 10 - 50 - 250 - 500 - 1000 KIU/L. (WHO 2 nd IRP 75/502). Ready to use.
Incubation Buffer: one vial (18 ml) containing TRIS buffer, pH 8.4, with 0.1% of
Tween 20 and 0.005% of Proclin 300 as preservative. Ready to use.
Sample Diluent: one vial (10 ml) of sample diluent consisting of horse serum, with
0.01% of Proclin 300 as preservative. Ready to use.
HRP-Streptavidin Conjugate: one vial (13 ml) of Streptavidin-Peroxidase
conjugate in a red colored buffer, pH 5.5 supplemented with 0.001% Proclin 300.
Ready to use.
A1002 – ALLERGEN Total IgE
M541 – Rev. 1 – 06/2010 – Pag. 11/32