A-070 A fully automated HbA1c Assay on the DxC 500 AU Clinical Chemistry Analyser

Abstract

Abstract Background Diabetes mellitus is a condition characterized by hyperglycemia resulting from the body’s inability to use blood glucose for energy. In Type-1 diabetes, the pancreas no longer makes insulin and blood glucose cannot enter the cells to be used for energy. In Type-2 diabetes, the pancreas does not make enough insulin or the body is unable to use insulin correctly. According to the World Health Organization (WHO), 422 million adults were living with diabetes globally in 2014 with an estimated 1.6 million deaths directly associated with diabetes annually. HbA1c is the major species of glycohemoglobin in human blood. HbA1c formation occurs through a non-enzymatic reaction, called glycation, which occurs between glucose and the n-terminal valine of the hemoglobin â-chain of HbA. A Schiff base intermediate product is formed, which rearranges to form a stable ketoamine in an irreversible reaction. The rate of glycation is proportional to the glucose concentration in the bloodstream and is an accurate reflection of average blood glucose over a period of eight to twelve weeks. HbA1c testing is recommended for the diagnosis of diabetes by the International Expert Committee (IEC), the American Diabetes Association (ADA), and the WHO, who recommended a diagnostic threshold of = 6.5% ( = 48 mmoL/moL) HbA1c and a range for pre-diabetes of 5.7%–6.4% (39–46 mmoL/moL) HbA1c. Methods The HbA1c Advanced assay on the DxC 500 AU* utilizes automatic sample pre-treatment, has batch and random access capability. No manual pre-treatment of the whole blood sample or additional washing steps are required. Firstly the red blood cells are hemolyzed automatically, total hemoglobin and glycated hemoglobin are then measured colorimetrically and immunoturbidimetrically respectively. Results Precision studies were conducted according to CLSI EP15-A3. Commercial controls and four native K2 EDTA whole blood samples ranging from 5.1% to 12.0% HbA1c, were run twice daily, over twenty days using three lots of reagent on three DxC500AU Clinical Chemistry analyzers at a single site. Repeatability ranged from 0.86% to 1.44% CV and Total Precision ranged from 1.55% to 2.43% CV. Linearity studies were conducted according to CLSI EP06-A, and verified an analytical measuring range of 4.0%–15.0% (NGSP) and 20–140 mmol/mol (IFCC). Method comparison and bias estimation was evaluated using CLSI EP09-A3. K2 EDTA patient samples (n = 150) across the analytical range were run vs a Secondary Reference method and yielded a slope of 1.031, intercept −0.147% HbA1c, correlation coefficient R = 0.997 for Weighted Deming regression, and slope 1.000 and intercept 0.06% HbA1c for Passing Bablok regression. Interference studies carried out demonstrated no significant interference from common endogenous interferences, a large panel of drugs, common Hb variants (HbC, HbD, HbE, HbA2 and HbS) and cross reactants (HbA0, HbA1a+b, acetylated hemoglobin, carbamylated hemoglobin, glycated hemoglobin, glycated albumin and labile HbA1c). Conclusion The HbA1c Advanced assay on the DxC 500 AU is a precise and accurate assay, requiring no manual pretreatment and can be used for monitoring and diagnosing diabetes. *Pending clearance by the United States Food and Drug Administration and achievement of CE compliance. Not currently available for in vitro diagnostic use.