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HbA1c IFU Biomax

The document provides detailed information on the quantitative determination of HbA1c in whole blood, emphasizing its significance as a long-term indicator of blood glucose levels in diabetic patients. It outlines the assay method, reagents used, potential interferences, and quality control measures, as well as expected values and performance characteristics. Additionally, it includes references for further reading on the topic.

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0% found this document useful (0 votes)
17 views2 pages

HbA1c IFU Biomax

The document provides detailed information on the quantitative determination of HbA1c in whole blood, emphasizing its significance as a long-term indicator of blood glucose levels in diabetic patients. It outlines the assay method, reagents used, potential interferences, and quality control measures, as well as expected values and performance characteristics. Additionally, it includes references for further reading on the topic.

Uploaded by

sharmashyamsingh
Copyright
© © All Rights Reserved
We take content rights seriously. If you suspect this is your content, claim it here.
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HbA1c .

INTENDED USE Mix and allow to stand for 5 minutes or until complete lysis is
Apparent
HbA1c is used for the quantitative determination of HbA1c in
whole blood. This reagent is for use on Fully automatic
INTERFERENCE AND LIMITATIONS
Biochemistry analyzer.
Due to its antibodies, HbA1c is a specific immunoassay for human
CLINICAL SIGNIFICANCE HbA1c. No interference was observed by ascorbic acid up to 60
Diabetes Mellitus is a chronic disease characterized by a mg/dl, conjugated and unconjugated bilirubin up to 40 mg/dl,
hyperglycemia. The consequences are metabolism disorders of lipemia up to 2000 mg/dl triglycerides, RF up to 250 IU/ml,
carbohydrates, lipids and proteins. The risk of complications carbamylated Hb up to 7.5 mmol/l, and acetylated Hb up to 5.0
associated with diabetes, including nephropathy, retinopathy mmol/l.
and cardiovascular diseases, increases in patients with poor No interference is observed by uremia, labile intermediates (Schiff
metabolic control. In the diabetic patients, where blood glucose base), and Hemoglobin variants HbS and HbA2. Elevated levels of
levels are elevated, HbA1c is formed as a consequence of the HbF may lead to falsely low HbA1c values. Alcoholism and ingestion
non-enzymatic -chain of haemoglobin molecule.glycation of of large doses of aspirin may lead to inconsistent results.
the N-terminus of the The level of HbA1c is proportional to the
level of glucose in the blood and has been widely accepted as WARNINGS AND PRECAUTIONS
an indicator of the mean daily blood glucose concentration over For in vitro diagnostic use.
the preceding 6-8 weeks. It is therefore, a long-term indicator of To be handled by entitled and professionally educated person
diabetic control, whereas, the measurement of blood glucose is Reagents of the kit are not classified like dangerous but contain
only a short-term indicator less than 0.1% sodium azide - classified as very toxic and
dangerous substance for the environment.
METHOD AND PRINCIPLE 2. Reagents S25: Avoid contact with eyes.
This reagent provides a quantitative assay for measuring 3. In very rare cases, samples of patients with gammopathy
concentrations of HbA1c in whole blood or red blood cell. It is might give falsified results.
based on latex agglutination. HbA1c test samples are 4. Immediately after HbA1c measurement cleaning of cuvettes
absorbed onto the surface of latex particles, which react with is necessary. Use the alkaline cuvette washing solution which is
Anti-HbA1c (antigen-antibody reaction). The turbidity caused recommended by the analyzer manufacturer.
by latex agglutination is measured at 660 nm, and the HbA1c 5. Take necessary precautions for use of laboratory reagent.
concentration in whole blood or red blood cell is calculated
from calibration curve. ASSAY PROCEDURE FULLY AUTO
1st reaction
REAGENT COMPOSITION Add 150 uL of R-1 Latex to 6 uL of the hemolyzed test sample
R1; Latex solution or each HbA1c calibrator, and after mixing the solution,
R2; Mouse anti-HbA1c monoclonal antibody, Goat Anti-mouse incubate it for five minutes at 37°C.
IgG 2nd reaction
Add 50 uL of R-2 solution and after mixing, incubate for five
Calibrator - Lyophilized. Store at 2 - 8 ºC. minutes at 37 °C.
Check the calibrator concentration on the bottle label. After Measure absorbance (turbidity) of each test sample and
reconstitution, the calibrator is stable for 7 days at 2 - 8 ºC and 30 respective HbAlc calibrators (at 660nm/800nm) according to
days at -20 ºC. the operating procedures for the autoanalyzer.
The calibration is valid only for the reagents and calibrator from the
same lot number. Reagents and calibrators are not Calculation of HbA1c concentration:
According to the operating procedures for the autoanalyzer,
interchangeable between kits with different lot numbers.
calculate the concentration of HbA1c in the test sample.
REAGENT PREPARATION When concentration (%) of test samples exceed the
Reagent R1 and R2 are ready to use as supplied. measurable range, dilute the test samples by 2-3 times with
HbA1c calibrator STD-2, or another test sample with a known
REAGENT STORAGE AND STABILITY concentration (%) of HbA1c. Then run the test again and
Unopened reagents, when stored at 2- 8 deg c temperature, and calculate the concentration (%) of HbA1c from the following
are stable up to the expiration date shown on the label. calculation formula;
Onboard stability and calibration frequency : HbA1c Concentration(%) of HbA1c
reagent has onboard stability of 3 weeks at 2-8 deg c. = [retested HbA1c (%) x n] - [HbA1c (%) of the sample/ a
Each laboratory has to define their own calibration calibrator used for the dilution x (n-1)]
frequency according to their workload and laboratory n = Dilution factor of the test sample (2-3).
conditions. [Calculation example for a test sample which exceeds the
measurable range]
REAGENT DETERIORATION When adding 200 uL of a known sample (HbA1c 4.5%) to 100
1. Avoid R-1 Latex from freezing or drying because it may uL of a test sample with approximately 17%, obtained by using
cause non-specific agglutination. Discard the reagent if it the test on the undiluted test sample. This means three times
does not meet stated performance parameters. dilution. If at retest, 9.1 % was obtained, the correct
concentration (%) of the test sample is calculated by following
the calculation formula;
SPECIMEN COLLECTION PREPARATION AND Concentration (%) of HbA1c = (9.1 x3)-(4.5x(3 - 1)) = 27.3 -
STORAGE 9.0 = 18.3 %
Whole blood collected with EDTA. Lysing procedure
below QUALITY CONTROL
Distilled water 500 μl It is recommended that controls be included in each set of
Whole blood Sample 10 μl assays. Commercially available control material with the
established Hba1c values may be used for quality control.
The assigned value of the control material must be
confirmed by the chosen application. Failure to obtain the
proper range of values in the assay of control material may
indicate reagent deterioration, instrument malfunction, or
procedural errors.
EXPECTED VALUES
4.6-6.2% (NGSP)
Reference intervals should be established or verified by the laboratory
based on an appropriate non-diabetic patient population.
%NGSP %IFCC mmol/mol
Non Diabetic 4-6 3-4 30-40
Target of Therapy <7 <5 <50
Change of >8 >6 >60
therapy
PERFORMANCE CHARACTERISTICS.
Measuring range: 3.3 to 16.0 %
Specificity: When 2 HbA1c control samples with a known
concentration (%) (high, middle, low) are tested according to
the test procedure described in the package insert, each test
result should be within ± 20% of respective known
concentrations (%) of the HbA1c control sample.
Sensitivity: When absorbance of HbA1c calibrator STD-
1(0%) is
measured according to the test procedure described in the
package
insert, the absorbance (turbidity) at 660 nm should be 0.50 or
less.
(2) When absorbance of HbA1c calibrator STD-5(15.6%) is
measured according to the test procedure described in the
package insert, the absorbance (turbidity) at 660 nm should
be 1.00 or more.
Method Comparison:
Study results in comparison with HPLC method.
n = 81
Correlation coefficient: r = 0.997
Regression equation: y = 0.979x + 0.0252

REFERENCES

1.Thomas L. Clinical Laboratory Diagnostics. 1st ed. Frankfurt:


TH-Books Verlagsgesellschaft; 1998. p. 142-48.
2. Sacks DB. Carbohydrates. In: Burtis CA, Ashwood ER,
editors. Tietz Textbook of Clinical Chemistry. 3rd ed.
Philadelphia: W.B. Saunders Company; 1999.p. 790-6.
3. Jeppsson JO, Kobold U, Barr J, Finke A et al. Approved
IFCC reference method for the measurement of HbA1c in
human blood. Clin Chem Lab Med 2002;40:78-89.
4. Hoelzel W, Weykamp C et al. IFCC Reference System for
Measurement of Hemoglobin A1c in Human Blood and the
National Standardization Schemes in the United States, Japan,
and Sweden: A Method-Comparison Study. Clin Chem 2004;
50:1:166-74.

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