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Effects of 7 Hemoglobin Variants on the Measurement of Glycohemoglobin by 14 Analytical Methods

¹ Department of Laboratory Medicine and Genetics, Samsung Medical Center, Sungkyunkwan University School of Medicine, Seoul, Korea; ² Department of Clinical Chemistry, Queen Beatrix Hospital, Winterswijk, The Netherlands; ³ Department of Laboratory Medicine and Genetics, Soonchunhyang University Bucheon Hospital and Soonchunhyang University College of Medicine, Gyeonggi-do, Korea;

^aaddress correspondence to this author at: Department of Laboratory Medicine and Genetics, Samsung Medical Center, Sungkyunkwan University School of Medicine, 50 Irwon-Dong, Gangnam-Gu, Seoul 135-710 Korea; fax 82-2-3410-2719, e-mail changski{at}skku.edu)

Abstract

Background: Hemoglobin variants (Hb^VAR) are not uncommon in the Korean population, with Hb G-Coushatta and Hb Queens being the 2 most common Hb^VAR. Hb G-Coushatta is also the most common Hb^VAR in Chinese people from the Silk Road region, as well as in some North American Indian tribes. However, data are scarce on the effect of these Hb^VAR on the different methods used for analyzing HbA_1c.

Methods: Specimens from 24 individuals with 7 Hb^VAR (Hb G-Coushatta, Hb Queens, Hb G-Hsi-Tsou, Hb Ube-4, Hb G-Waimanalo, Hb Inglewood, and Hb Bologna-St.Orsola) were collected and tested using the International Federation of Clinical Chemistry primary reference method as well as 14 routine HbA_1c assay methods.

Results: Hb G-Coushatta showed a clinically significant effect on the measured HbA_1c, particularly when analysis was performed with ion-exchange HPLC methods with short elution times. This interference could be resolved by measuring the HbA_1c using other methods such as HPLC with a long elution time, immunoassay, boronate affinity chromatography, and enzymatic assay. Hb Queens showed a clinically significant difference, defined as a >10% deviation from regression lines, in results from the 2 HPLC methods but not in the other methods. The remaining 5 rare Hb^VAR showed different HbA_1c results in the different assays.

Conclusion: Hb G-Coushatta, Hb Queens, and other rare Hb^VAR can interfere with glycohemoglobin assays, including ion-exchange HPLC methods with short elution times, but the interference can be resolved using other unaffected methods. It is important to identify these Hb^VAR through a careful inspection of the chromatograms and apply other noninterfering methods for accurate measurements of the HbA_1c.

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