Objectives:Hemoglobin (Hb) variants remain an important cause of erroneous HbA1c results. We present an approach to overcome the interference of Hb variants on HbA1c measurements using matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS). Methods:Samples containing or not containing Hb variants were analyzed for HbA1c using an MALDI-TOF MS system (QuanTOF) and a boronate affinity comparative method (Ultra2). For QuanTOF, two sets of HbA1c values were obtained through α- and β-chain glycation. Results:A robust correlation between the glycation degrees of the α- and β-chains was found, and HbA1c values derived from α- and β-chain glycation correlated well with the Ultra2 results. Statistically significant differences (p<0.01) were found for all the Hb variants tested. When using the conventional β-chain glycation to determine HbA1c, clinically significant differences were only found among samples containing β-chain variants detected by QuanTOF (i.e., Hb J-Bangkok, Hb G-Coushatta, and Hb G-Taipei). In contrast, based on α-chain glycation, no clinically significant differences were found for these three variants. Conclusions:In addition to conventional β-chain glycation, α-chain glycation can be used to calculate HbA1c values. The interference of Hb variants on HbA1c quantification can be overcome by employing the glycation of the globin chain without a genetic variant to estimate HbA1c values.

译文

目的:血红蛋白(Hb)变异仍然是导致错误的HbA1c结果的重要原因。我们提出了一种使用基质辅助激光解吸/电离飞行时间质谱仪(MALDI-TOF MS)来克服Hb变体对HbA1c测量的干扰的方法。
方法:使用MALDI-TOF MS系统(QuanTOF)和硼酸酯亲和比较方法(Ultra2)对包含或不包含Hb变体的样品进行HbA1c分析。对于QuanTOF,通过α和β链糖基化获得了两组HbA1c值。
结果:发现α链和β链的糖基化程度之间存在强相关性,并且α链和β链糖基化得到的HbA1c值与Ultra2结果具有很好的相关性。在所有测试的Hb变体中发现统计学上的显着差异(p <0.01)。当使用常规的β链糖基化来确定HbA1c时,仅在包含通过QuanTOF检测到的β链变体的样品(即Hb J-Bangkok,Hb G-Coushatta和Hb G-Taipei)中发现临床上的显着差异。相反,基于α链糖基化,这三个变体没有发现临床上的显着差异。
结论:除常规的β链糖基化外,α链糖基化还可用于计算HbA1c值。 Hb变体对HbA1c定量的干扰可以通过不带遗传变体的球蛋白链糖基化来估计HbA1c值来克服。

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