i-CHROMATM HbA1c (Pipetting Method) - Al-Sakr Al-Waheed ...

BODITECH MED INC. i-CHROMA HbA1c 25
REFERENCES
1. Goldstein DE, Little RR, Lorenz RA, Malone JI, Nathan D,
Peterson CM. Tests of glycemia in diabetes. Diabetes Care 1995;
18:896-909.
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diabetes. Am J Med 1981; 70:325-30.
3. Jovanovic L, Peterson CM. The clinical utility of glycosylated
hemoglobin. Am J Med 1981; 70:331-8.
4. Nathan DM, Singer DE, Hurxthal K, Goodson JD. The clinical
information value of the glycosylated hemoglobin assay. N Engl
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5. Goldstein DE, Little RR, Wiedmeyer HM, England JD,
McKenzie EM. Glycated hemoglobin: methodologies and
clinical applications. Clin. Chem. 1986; 32:B64-70.
6. Goldstein DE, Little RR, England JD, Wiedemeyer H-M,
McKenzie E. Methods of glycosylated hemoglobins: high
performance liquid chromatography and thiobarbituric acid
colorimetric methods. In: Clarke WL, Larner J, Pohl SL, eds.
Methods in diabetes research, Vol. 2. New York: John Wiley,
1986:475-504.
7. Tahara Y, Shima K. The response of GHb to stepwise plasma
glucose change over time in diabetic patients. Diabetes Care
1993; 16:1313-4.
8. Svendsen PA, Lauritzen T, Soegaard U, Nerup J. Glycosylated
haemoglobin and steady-state mean blood glucose concentration
in type 1 (insulin-dependent) diabetes. Diabetologia 1982;
23:403-5.
9. Cefalu WT, Wang ZQ, Bell-Farrow A, Kiger FD, Izlar C.
Glycohemoglobin measured by automated affinity HPLC
correlates with both short-term and long-term antecedent
glycemia. Clin Chem 1994; 40:1317-21.
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glycemia in diabetes mellitus. Their use in establishing a
diagnosis and in treatment. Ann Intern Med 1989; 110:125-37.
11. Molnar GD. Clinical evaluation of metabolic control in
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Rev. 11_100125_Multi
i-CHROMA TM HbA1c
(Pipetting Method)
ImmunoAssay for Quantitative Measurement of
Hemoglobin A1c in Human blood with i-CHROMA TM
Reader System
INTENDED USE
The i-CHROMA TM HbA1c Test along with i-CHROMA TM Reader
is a fluorescence immunoassay that measures a ratio of hemoglobin
A1c to total hemoglobin in whole blood.
The test is used for routine monitoring of long-term glycemic status
in patients with diabetes mellitus.
INTRODUCTION
Glycated proteins are formed post-translationally from the slow,
nonenzymatic reaction between glucose and amino groups on
proteins 1 . For hemoglobin, the rate of synthesis of Hemoglobin A1c
(HbA1c) is principally a function of the concentration of glucose to
which the erythrocytes are exposed. HbA1c is a clinically useful
index of mean glycemia during the preceding 120 days, the average
life span of erythrocytes 1, 2-9 . Although carefully controlled studies
have documented a close relationship between the concentrations
of HbA1c and mean glycemia, routine determinations of blood
glucose by patients or by their healthcare providers are not
considered as reliable as HbA1c to quantify mean glycemia 1, 3, 4, 10-12 .
Concentrations of other blood-based glycated proteins (e.g.,
glycated serum/plasma proteins, "fructosamine") also reflect mean
glycemia, but over a much shorter time than HbA1c: 15-30 days
and 60-120 days, respectively 1, 2-10, 13, 14 .
PRINCIPLE
The i-CHROMA TM HbA1c Test is based on fluorescence
immunoassay technology 15 . The i-CHROMA TM HbA1c Test uses
a competitive immunodetection method, such that by mixing
detector buffer with hemolysed whole blood in test vial, the
fluorescence-labeled detector anti-HbA1c antibody in buffer binds
to HbA1c in blood specimen. As the sample mixture is loaded onto
the sample well of the test device and migrates the nitrocellulose
matrix of test strip by capillary action, the detector antibody
competitively binds to HbA1c in the sample and glycated
hemoglobin that has been immobilized on test strip. Thus as the
more HbA1c antigen is in blood specimen, signal intensity of
fluorescence of detector antibody decreases proportionally. The
default result unit of i-CHROMA TM HbA1c Test is displayed as a
percentage(%) from i-CHROMA TM Reader. The working range of
i-CHROMA TM HbA1c Test system is 4–15%.
* Reference Value : < 6.5 %
COMPOSITION OF REAGENTS

BODITECH MED INC. i-CHROMA HbA1c 25
The i-CHROMA TM HbA1c Test Kit consists of Test Device,
Blood Collecting Capillary, and Detector Buffer tube. Test Device
is individually sealed with a desiccant in aluminum pouch. Detector
Buffer/Hemolysis buffer is pre-dispensed individually in a tube,
packed in a box, and delivered separately from Test Device in a
styrofoam box filled with ice pack.
• Test Device contains a test strip in which glycated hemoglobin
and avidin have been immobilized on the test and on the control
line of strip, respectively.
• Hemolysis buffer is pre-dispensed individually in a small tubes
and composed of cationic detergent
• Detector Buffer is dispensed in a vial. It contains fluorescencelabeled
goat anti-HbA1c (mouse monoclonal, 4.5µg/mL),
fluorescence-labeled BSA-biotin (200 ng/mL), 1% gelatin as a
stabilizer, and 0.1% Sodium Azide as a preservative in PBS.
WARNINGS AND PRECAUTIONS
• IVD For In Vitro Diagnostic Use.
• Carefully follow the instructions and procedures described in this
insert. REF Catalog No. HbA1c -25T
• Don’t use Test Device if its lot number does not match with ID
Chip that is inserted onto the instrument.
• The i-CHROMA TM HbA1c Test Kit is only operational in the i-
CHROMA TM Reader. And tests should be applied by trained staff
working in the laboratories where the sample(s) is taken by
qualified medical personnel.
• LOT Neither inter-change materials from different product lots
nor use beyond the expiration date. The use of medical
device beyond expiration date may affect on test result.
• The i-CHROMA TM HbA1c Test Device should remain in its
original sealed pouch until ready to use. Do not use the Test
Device if the pouch is damaged or the seal is broken. Discard
after single use.
• The i-CHROMA TM HbA1c Test Device and Reader should be
used away from vibration and magnetic field. During normal
usage, i-CHROMA TM HbA1c Test may introduce minute
vibration, which should be regarded normal.
• Use separate clean pipette tips and sample vials for different
specimens. The pipette tips and sample vials should be used
for one specimen only. Discard after single use.
• Blood specimens, used test devices, pipette tips and sample vials
are potentially infectious. Proper laboratory safety
techniques, handling and disposal methods should be
followed in accordance with standard procedures and
relevant regulations observed by microbiological hazard
materials..
• The test will be applied on a routine basis and not in emergency
situations.
• Do not smoke, eat, or drink in areas in which specimens or kit
reagents are handled.
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STROAGE AND STABILITY
• Store the Hemolysis and Detector buffer in a refrigerator at 2° -
8°C. The Hemolysis and Detector Buffer are stable up to
20 months.
• Once removed from refrigerator, allow the Hemolysis and
Detector Buffer for 30 minutes to return to room temperature
before testing.
• Store i-CHROMA TM HbA1c Test Device at 4-30°C in its sealed
box. The i-CHROMA TM HbA1c Test Device is stable for
20 months (while in the sealed pouch) if stored at 4-30°C.
• If stored in a refrigerator, allow a minimum of 30 minutes for the
Test Device to reach room temperature while it is in the sealed
pouch.
• Do not remove the device from the pouch until ready to use. The
Test Device should be used immediately once opened.
• The storage and shipping of Test Kit should be complied as
indicated in manual. However, it is remotely possible that only
part of test Kit is affected by stability problems.
SAMPLE COLLECTION AND PREPARATION
The test can be performed with human blood samples.
• The whole blood specimen must be at room temperature and be
homogeneous before testing. Fresh blood samples must be used a
immediately as possible, within 24 hours after collection. .
MATERIALS PROVIDED
BodiTech Med Incorporated i-CHROMA TM HbA1c Test
Catalog REF Catalog No. HbA1c-25T
Kit contains:
Test Devices 25T/box
Hemolysis Buffer 25 tubes/box
Detection buffer 1 vial
MATERIALS REQUIRED BUT NOT PROVIDED
i-CHROMA TM Reader REF Catalog No. FR-203
Thermal Printer
Transfer pipette
Glass capillary (5 ul)

BODITECH MED INC. i-CHROMA HbA1c 25
PROCEDURE
• Image of the test kit
1. Set a Test Device on a dust-free clean place.
2. Check/insert ID chip onto an instrument. Make sure
that the Test Device lot # matches with ID chip #.
3. Take out a Detector Buffer vial and Hemolysis buffer tubes
from a refrigerator and leave it at room temperature.
4. Take 100 ul of Detection Buffer from the vial, put into the
pre-dispensed Hemolysis buffer tube, and mix well.
5. Make a prick on a fingertip or prepare a tube blood.
6. Draw 5 µL of whole blood with a glass capillary.
7. Put the glass capillary into the Hemolysis buffer tube and
shake the tube 10 times by inversion to take the blood out
of the capillary.
8. Apply 50 ul of the mixture onto the sample well and 100 ul
onto the Hb sample well of the test device cartridge (refer
the picture above)
9. Leave the Test Device at room temperature for 12 min before
inserting the device into the holder.
10. To start scanning, insert test device onto the holder of
i-CHROMA TM Reader and press “SELECT” button.. Make
sure direction of Test Device and push the device back all
the way.
11. The instrument will automatically start to scan the Test
Device immediately.
12. Read the results on the display screen of i-
CHROMA TM Reader or tear out the print for record.
� Refer to i-CHROMA TM Reader Operation Manual for the
complete instructions on use of the Test.
RESULT
REF Catalog No. FR- 203
The i-CHROMA TM Reader calculates HbA1c test results
automatically and displays percentage (%) of Hemoglobin A1c in
blood on the LCD.
For further information, refer to the Operation Manual for the i-
CHROMA TM Reader.
Rev. 11_100125_Multi
Quality Control
� A quality control test using commercially available controls
should be performed as a part of good testing practice, to
confirm the expected QC results, to confirm the validity of the
assay, and to assure the accuracy of patient results. If you want
to perform QC of Test Kit, we recommend using Boditech
Med’s i-CHROMA HbA1c control.
� A quality control test should be performed at regular intervals,
and before using a new kit with patient specimens, controls
should be tested to confirm the test procedure, and to verify the
tests produce the expected QC results. QC specimens should also
be run whenever there is any question concerning the validity of
results obtained. Upon confirmation of the expected results, the
test device is ready to use with patient specimens. Control
standards are not provided with this test kit. For information
about obtaining the controls, contact BodiTech Med Inc.
Technical Services for assistance.
Procedure Control
� Each i-CHROMA TM HbA1c Test Device contains internal
control that satisfies routine quality control requirements. This
internal control is performed each time a patient sample is tested.
This control indicates that the test device was inserted and read
properly by i-CHROMA TM Reader. An invalid result from the
internal control causes an error message on i-CHROMA TM
Reader indicating that the test should be repeated.
LIMITATIONS OF THE PROCEDURE
• The results of i-CHROMA TM HbA1c Test should be evaluated
with all clinical and laboratory data available. If HbA1c Test
results do not agree with the clinical evaluation, additional tests
should be performed.
• The false positive results include cross-reactions with some
components of blood from individual to antibodies; and nonspecific
adhesion of some components in human blood that have
similar epitopes to capture and detector antibodies. In the case of
false negative results, the most common factors are: nonresponsiveness
of antigen to the antibodies by that certain
unknown components are masking its epitope, such that antigen
cannot be seen by the antibodies; instability of A1c antigen,
resulting in degradation with time and, or temperature, such that
they become no longer recognizable by antibodies; and degraded
other test components. The effectiveness of the test is highly
dependent on storage of kits and sample specimens at optimal
conditions.
• Other factors may interfere with i-CHROMA TM HbA1c Test and
may cause erroneous results. These include technical or
procedural errors, as well as additional substances in blood
specimens.
PERFORMANCE CHARACTERISTICS
1. Analytical Sensitivity: Analytical sensitivity means the lowest
concentration of HbA1c that the test system can detect with
CV

BODITECH MED INC. i-CHROMA HbA1c 25
2. Specificity: Other bio-molecules, such as Bilirubin, Triglyceride,
Ascorbic acid and glucose were added to test specimen with
much higher level than their physiological level in normal blood.
There was no significant interference with the HbA1c
measurement, nor was there any significant assay cross-reactivity
with other disease-related biomarkers in blood.
3. Imprecision: For the intra-assay imprecision, 20 replicates were
tested at each control sample. For the imprecision evaluation,
tests were conducted on 10 sequential days with 5 replicates and
for 3 persons at each HbA1c concentration.
HbA1c (%)
Imprecision of i-CHROMA TM HbA1c
Intra-assay Inter-assay
Mean S.D CV% Mean S.D CV%
5.6 5.4 0.3 5.6 5.5 0.26 5.0
10.1 9.7 0.41 4.3 10.2 0.42 4.1
4. Linearity: The coefficient of linear regression was R=0.997.
Linearity of i-CHROMA TM HbA1c Test was 4~15%.
5. Comparability: The HbA1c concentrations of 152 clinical
specimens were quantified independently with i-CHROMA TM
HbA1c and Bio Rad Variant II(HPLC) automatic analyzer. The
test results were compared and their compatibilities were
investigated with linear regression and correlation of coefficient
(R). i-CHROMA TM HbA1c was comparable well to other
method (R=0.97).
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