Clinical Chemistry Siemens Point of Care - Urinalysis
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Clinical Chemistry 49: 269-274, 2003; 10.1373/49.2.269
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(Clinical Chemistry. 2003;49:269-274.)
© 2003 American Association for Clinical Chemistry, Inc.

New Enzymatic Assay for Glycohemoglobin

Ikunosuke Sakurabayashi1, Tatsurou Watano1, Satoshi Yonehara2a, Kaori Ishimaru2, Kaoru Hirai2, Tsuguki Komori2 and Masayuki Yagi2

1 Department of Clinical Pathology, Jichi Medical School Omiya Medical Center, 1-847 Amanuma-cho, Saitama-shi, Saitama-ken, Japan 330-8503.

2 ARKRAY, Inc., 57 Nishi Aketa-Cho, Higashi-Kujo Minami-Ku, Kyoto, Japan 601-8045.

aAuthor for correspondence. Fax 81-75-661-9435; e-mail yonehara{at}arkray.co.jp.

Background: Previous methods to measure glycohemoglobin (GHb) have been time-consuming or imprecise; we therefore developed a new enzymatic assay for GHb.

Methods: Blood cells were first hemolyzed, and hemoglobin was digested with protease to yield fructosyl amino acid. Fructosyl amino acid oxidase acts on the fructosyl amino acid and generates hydrogen peroxide, which reacts with chromogens in the presence of peroxidase. Total hemoglobin was measured spectrometrically in the same reaction tube. The results were reported as the ratio of the concentrations of GHb and hemoglobin.

Results: The measured values were comparable to those determined with a HPLC method and with an immunoassay in blood samples from 2854 patients with diabetes. Regression analysis for the enzymatic assay (y) vs the HPLC method (x) produced the following: r = 0.979; slope, 0.994 [95% confidence interval (CI), 0.986–1.001]; y-intercept, 0.04% (95% CI, -0.09% to 0.01%); n = 2854. For the enzymatic assay (y) vs the immunoassay (x), the regression statistics were as follows: r = 0.982; slope, 1.002 (95% CI, 0.995–1.009); y-intercept, 0% (95% CI, -0.05% to 0.05%); n = 2854.

Conclusions: The values measured by the new enzymatic assay are sufficiently correlated with those of the conventional HPLC method and immunoassay, but the proposed assay for GHb is rapid and has high precision.






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Copyright © 2003 by the American Association for Clinical Chemistry.