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Comparison of three methods for measurement of blood hba1c as to reliability

Boinapalli Sudhakar*, Aluguballi Sreenivas Reddy, J. J.J. Fallerio


To compare three methods for measurement of blood HbA1c as to reliability, ease and time consumption. Measurements of HbA1c were made in blood from 230 patients with pre-diabetes and diabetes using a turbidimetric inhibition immunoassay (TINIA), which also required measurement of total hemoglobin, a particle-enhanced immune turbidimetric assay (PEITT) without measurement of total hemoglobin, and high performance liquid chromatography (HPLC). There was good concordance between results of PEITT and HPLC methods (r = 0.9401, p<0.0001 and by Deming Regression; y = 0.9978x + 0.24). The average HbA1c (7.52±1.40 %) measured by HPLC was higher than the other methods (TINIA: 7.12±1.66 % and PEITT: 7.26±1.39 %, p<0.0001). The measured total time spent on 240 samples was 81 min. for TINIA, 54min. for PEITT and 540 min. for HPLC. It has been found that, the PEITT method, which is reliable, faster, and easier to perform, can be used as an alternative to TINIA and HPLC measuring system within the known imprecision limits.”


HbA1c; Particle Enhanced Immunoturbidimetric Test; Turbidimetric Immunoassay; HPLC

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The Diabetes Control and Complications Trial Research Group. The effect of intensive treatment of diabetes on the development and progression of long-term complications in insulin dependent diabetes mellitus. N Engl J Med. 1993. 14: 977-986.

Krishnamurti U, Steffes MW. Glycohemoglobin: a primary predictor of the development or reversal of complications of diabetes mellitus, clinical chem., 2001, 47, 1157-65.

American Diabetes Association Clinical Practice Recommendations. Diabetes Care. 2001. 24 Suppl. 1: S1-133.

Nuttall FQ. Comparison of percent total GHb with percent HbA1c in people with and without known diabetes. Diabetes Care, 1998, 21, 1475-1480.

Baynes JW, Bunn HF, Goldstein D, Harris M, Martin DB, Peterson C, Winter halter K. National Diabetes Data Group: report of the expert committee on glucosylated hemoglobin. Diabetes Care, 1984, 7, 602-606.

Nathan DM, Singer DE, Hurxthal K, Goodson JD. The clinical information value of the glycosylated hemoglobin assay. N Engl J Med 1984. 310: 341-346.

Hoelzel W, Weykamp C, Jeppsson JO, Miedema K, Barr JR, Goodall I, Hoshino T, John WG, Kobold U, Little R, Mosca A, Mauri P, Paroni R, Susanto F, Takei I, Thienpont L, UmemotoM, Wiedmeyer HM. 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, 166-174.

Santiago JV. Lessons from the Diabetes Control and Complications Trial. Diabetes. 1993; 42: 1549-1554.

Weykamp CW, Penders TJ, Muskiet FA, van der Slik W. Influence of hemoglobin variants and derivatives on glycohemoglobin determinations, as investigated by 102 laboratories using16 methods. Clin Chem, 1993, 39, 1717-1723.

Weykamp CW, Penders TJ, Siebelder CW, Muskiet FA, van derSlik W, Interference of carbamylated and acetylated hemoglobinsin assays of glycohemoglobin by HPLC, electrophoresis, affinity chromatography, and enzyme immunoassay. ClinChem, 1993, 39, 138-142.

Little RR, Tennill AL, Rohlfing C, Wiedmeyer HM, KhannaR, Goel S, Agrawal A, Madsen R, Goldstein DE. Can glycohemoglobin be used to assess glycemic control in patients with chronic renal failure? Clin Chem, 2002, 48,784-786.

Metus P, Ruzzante N, Bonvicini P, Meneghetti M, ZaninottoM, Plebani M. Immuno turbidimetric assay ofglycated hemoglobin. J Clin Lab Anal, 1999, 13, 5-8.

Chang J, Hoke C, Ettinger B, Penerian G. Evaluationand interference study of hemoglobin A1c measured by turbidimetric inhibition immunoassay. Am J Clin Pathol, 1998,109, 274-278.

Goodall I. HbA1c standardisation destination globalI FCC Standardisation. How, why, where and when--a tortuous pathway from kit manufacturers, via inter-laboratory lyophilizedand whole blood comparisons to designated national comparison schemes. Clin Biochem Rev, 2005. 26, 5-19.

Davie SJ, Gould BJ, Yudkin JS. Effect of vitamin Con glycosylation of proteins. Diabetes 41 (2): 167-173.

Rohlfing CL, Connolly SM, England JD, Hanson SE, Moellering CM, Bachelder JR, Little RR. (2008) The effect ofelevated fetal hemoglobin on hemoglobin A1c results: five commonhemoglobin A1c methods compared with the IFCC reference method. Am J Clin Pathol, 1992, 129, 811-814.

Little RR, England JD, Wiedmeyer HM, Goldstein DE, Effects of whole blood storage on results for glycosylated hemoglobin as measured by ion-exchange chromatography, affinity chromatography, and colorimetry. Clin Chem, 1983, 29, 1113-1115.

Peterson CM, Jovanovic L, Raskin P, Goldstein DE. Acomparative evaluation of glycosylated haemoglobin assays: feasibility of references and standards. Diabetologi, 1984, 26, 214-217.

Geistanger A, Arends S, Berding C, Hoshino T, Jeppsson JO, Little R, Siebelder C, Weykamp C. Statistical methods for monitoring the relationship between the IFCC reference measurement procedure for hemoglobin A1c and the designated comparison methods in the United States, Japan, and Sweden, Clin Chem, 2008, 54, 1379-1385.

Sacks DB, Bruns DE, Goldstein DE, Maclaren NK, Mc Donald JM, Parrott M. Guidelines and recommendations for laboratory analysis in the diagnosis and management of diabetes mellitus, Clin Chem, 2002, 48, 436-472.

Weykamp CW, Penders TJ, Muskiet FA, van der Slik W. Effect of calibration on dispersion of glycohemoglobin values determined by 111 laboratories using 21 methods. Clin Chem, 1994, 40, 138-144.

Weykamp CW, Penders TJ, Miedema K, Muskiet FA, van der Slik W. Standardization of glycohemoglobin results and reference values in whole blood studied in 103 laboratories using20 methods. Clin Chem, 1995, 41, 82-86.



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