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Clinical Chemistry and Laboratory Medicine (CCLM)

Published in Association with the European Federation of Clinical Chemistry and Laboratory Medicine (EFLM)

Editor-in-Chief: Plebani, Mario

Ed. by Gillery, Philippe / Greaves, Ronda / Lackner, Karl J. / Lippi, Giuseppe / Melichar, Bohuslav / Payne, Deborah A. / Schlattmann, Peter

IMPACT FACTOR 2017: 3.556

CiteScore 2018: 2.44

SCImago Journal Rank (SJR) 2018: 1.191
Source Normalized Impact per Paper (SNIP) 2018: 1.205

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Volume 49, Issue 7


Adaptation of the Diazyme Direct Enzymatic HbA1c Assay for a microplate reader at room temperature

Ilkka Penttilä / Karri Penttilä / Toivo Halonen / Kari Pulkki / Jukka Törrönen / Rainer Rauramaa
Published Online: 2011-05-17 | DOI: https://doi.org/10.1515/CCLM.2011.186


Background: Measurement of hemoglobin A1c (HbA1c) is a key diagnostic criterion and a key parameter for the follow-up of the treatment of diabetes mellitus. Typically, immunochemical assays of HbA1c are performed in clinical chemistry analyzers. In this study, we applied the HbA1c assay on a microplate reader at room temperature.

Methods: HbA1c samples were measured using the Direct Enzymatic HbA1c Assay™ from Diazyme Laboratories (Poway, CA, USA) using a Plate Chameleon™ Microplate Reader (Hidex Co., Turku, Finland) according to the manufacturer's protocol and a modification of the method to room temperature. The Tosoh G7 HPLC method for HbA1c (Tosoh Co., Tokyo, Japan) was used as a comparative method.

Results: There was good correlation of HbA1c results when the assay was performed at room temperature (+22°C) compared with that at +37°C (r=0.987). The modified method was linear over the HbA1c range 4%–14%. Analysis of HbA1c results from 50 blood samples by the modified method showed good agreement with the HPLC method (r=0.990).

Conclusions: The modified Diazyme Direct Enzymatic HbA1c Assay™ appears to work as good at +22°C as that performed according to manufacturer's protocol at +37°C.

Keywords: diabetes; hemoglobin A1c; immunoassay; plate reader photometer

About the article

Corresponding author: Ilkka Penttilä, MD, PhD, Kuopio Research Institute of Exercise Medicine, Haapaniementie 16, 70100 Kuopio, Finland

Received: 2010-11-18

Accepted: 2011-02-06

Published Online: 2011-05-17

Published in Print: 2011-07-01

Citation Information: Clinical Chemistry and Laboratory Medicine, Volume 49, Issue 7, Pages 1221–1223, ISSN (Online) 1437-4331, ISSN (Print) 1434-6621, DOI: https://doi.org/10.1515/CCLM.2011.186.

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