Ye Dan, Tang Yan, Zhang Mei
Department of Laboratory Medicine, West China Hospital, Sichuan University, Chengdu 610041, China.
Department of Laboratory Medicine, The First People's Hospital of Longquanyi District, Chengdu 610100, China.
Sichuan Da Xue Xue Bao Yi Xue Ban. 2023 Sep;54(5):1019-1023. doi: 10.12182/20230960210.
To examine the common types of hemoglobin variants and to evaluate the influence of common variants on the results of two kinds of glycosylated hemoglobin (HbA1c) tests.
We conducted a retrospective study, analyzing the data of a patient population undergoing two HbA1c tests, high performance liquid chromatography (HPLC) and capillary electrophoresis (CE), at West China Hospital, Sichuan University between March 2021 and February 2022. By screening the chromatograms, the hemoglobin variants were identified and their migration positions in the CE method were recorded. The effects of the variants with different migration positions on the findings of the two methods were compared. Variant samples with different migration positions were selected and Sanger sequencing was performed to determine mutations in HBA1, HBA2, and HBB genes in the variant samples.
We examined the HbA1c of 207 786 patient samples, identifying variant peaks in the chromatograms of 372 patients. The detection rate of variants was 0.18%, with the variant identification rate of HPLC being 43.3% and that of CE, 100%. Through sequencing, 20 variants were detected. A total of 261 patient samples were tested for HbA1c with both HPLC and CE. HPLC reported all HbA1c results, while CE did not report HbA1c results for 28 samples, among which, 26 showed abnormal peaks that overlapped with HbA1c peaks, and 2 showed abnormal peaks that overlapped with HbA0 peaks. The commonly observed variant migration positions, as revealed by CE, were at the horizontal coordinates of 225±1, 200±3, 100±2, 124±1, 70±2, and 182±1. There was significant difference between HPLC method and CE method in the determination of HbA1c ( <0.0083), and the difference between the two methods was the largest when there were variants in the 200±3 region. Linear regression showed that the correlation of HbA1c results between the two methods was different when different regional variants were present, and that the correlation between the two methods was strongest when 124±1 region was present ( =0.998).
There are diverse types of hemoglobin variants and most of them can affect the HbA1c findings of HPLC. Analyzing the chromatogram facilitates the identification of the variants.
研究常见血红蛋白变异体类型，并评估常见变异体对两种糖化血红蛋白（HbA1c）检测结果的影响。
我们进行了一项回顾性研究，分析了2021年3月至2022年2月期间在四川大学华西医院接受两种HbA1c检测（高效液相色谱法（HPLC）和毛细管电泳法（CE））的患者群体数据。通过筛查色谱图，识别血红蛋白变异体并记录其在CE法中的迁移位置。比较不同迁移位置的变异体对两种方法检测结果的影响。选择不同迁移位置的变异体样本进行Sanger测序，以确定变异体样本中HBA1、HBA2和HBB基因的突变情况。
我们检测了207786例患者样本的HbA1c，在372例患者的色谱图中识别出变异峰。变异体的检出率为0.18%，HPLC的变异体识别率为43.3%，CE的变异体识别率为100%。通过测序，检测到20种变异体。共有261例患者样本同时接受了HPLC和CE的HbA1c检测。HPLC报告了所有HbA1c结果，而CE未报告28例样本的HbA1c结果，其中26例显示异常峰与HbA1c峰重叠，2例显示异常峰与HbA0峰重叠。CE显示的常见变异体迁移位置的横坐标为225±1、200±3、100±2、124±1、70±2和182±1。HPLC法和CE法在HbA1c测定方面存在显著差异（<0.0083），当200±3区域存在变异体时，两种方法之间的差异最大。线性回归显示，当存在不同区域变异体时，两种方法的HbA1c结果相关性不同，当存在124±1区域变异体时，两种方法之间的相关性最强（=0.998）。
存在多种类型的血红蛋白变异体，其中大多数会影响HPLC的HbA1c检测结果。分析色谱图有助于识别变异体。
