Glycated Hemoglobin A1c (HbA1c) Assay Kit
Method: Enzymatic Hemoglobin A1c (HbA1c) assay
Wavelength: 570nm, 700nm
Linearity range: 4% - 15%
Intended Use
Glycated Hemoglobin A1c (glycated hemoglobin A1c; A1c; HbA1c) assay kit reagents are intended for use in the quantitative determination of stable HbA1c in human whole blood samples. HbA1c is expressed as a percentage of the total hemoglobin (THb). For in-vitro diagnostic use.
Introduction
Glycated Hemoglobin A1c (HbA1c) is an important test recommended by the American Diabetes Association (ADA) and its usefulness was clarified by the United Kingdom Prospective Diabetes Study (UKPDS) and Diabetes Control and Clinical Trials (DCCT). Currently, the HbA1c test is recommended for patients with diabetes every 2-3 months as part of the patient Diabetes management program. Glycohemoglobin is produced by non-enzymatic addition of glucose to amino groups in hemoglobin. HBA1c refers to glucose modified hemoglobin A (HbA) specifically at N-terminal valine residues of hemoglobin beta chains. HbA1c test used both as an index of mean glycemia and as a measure of risk for the development of diabetes complications1-3. Therefore, the A1c test is a good indicator of glycemic control in the preceding 2-3 months.
Assay Principle
Glycated Hemoglobin A1c (HbA1c) assay kit is an enzymatic assay in which lysed whole blood samples are subjected to extensive protease digestion with Bacillus sp protease. This process releases amino acids including glycated valines from the hemoglobin beta chains. Glycated valines then serve as a substrate for recombinant fructosyl valine oxidase (FVO) enzyme, produced in E. coli. FVO specifically cleaves N-terminal valines and produces hydrogen peroxide. This, in turn, is measured using a horse radish peroxidase (POD) catalyzed reaction and a suitable chromagen. Total hemoglobin is determined separately by conversion of all hemoglobin derivatives of the samples into hematin using an alkaline method as described in Zander et al (1984).
Fresh whole blood samples are treated with lysis reagent to release hemoglobin. The same lysate is then subjected to two parallel tests. The first determines glycated hemoglobin (GHb) content and, the second test determines total hemoglobin (THb) content of the samples. The HbA1c concentration is expressed as a concentration ratio of glycated hemoglobin to total hemoglobin.
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