多重PCR筛选K、Yt^b血型抗原的方法研究

Development of a multiplex polymerase chain reaction system for screening low-frequency blood group antigens K and Yt^b

目的建立筛选稀有血型抗原K、Yt^b的多重PCR体系，了解这两种血型抗原在维吾尔族人群中的分布情况。方法针对血型抗原K、Yt^b的编码基因KEL和ACHE单核苷酸多态性位点设计序列特异性引物，通过优化PCR反应条件，构建可同时检测K、Yt^b血型抗原的多重PCR体系，用该体系对362份新疆维吾尔族随机献血者样本进行相应抗原筛选，并对筛选到的稀有样本进行杂合性分析。结果成功构建可同时检测低频血型抗原K、Yt^b的多重PCR体系，362份维吾尔族样本中共检出9例K＋k＋，41例Yt（a＋b＋），4例Yt（a—b＋）。结论建立的检测K、Yt^b血型抗原的多重PCR体系简便有效。通过筛选所获得的维吾尔族稀有血型数据，可为临床输注配合性血液提供参考资料。

Objective A multiplex PCR system for screening rare blood group antigens K and Yt^b was constructed to study the distribution of the two blood groups in a Uygur population in Xinjiang, China. Methods Sequence-specific primers （SSP） were designed based on single nucleotide polymorphism sites of KEL and ACHE alleles encoding the two blood group antigens. The system was designed for simultaneously detecting the two antigens by optimizing the PCR reaction. Three hundred and sixty-two randomly selected healthy individuals were screened. Products of PCR were further analyzed for heterozygosity. Results The system was set up successfully. No KK sample was identified and 9 K＋k＋, 41 Yt （a＋b＋）, 4 Yt （a-b ＋） were found among the 362 samples. Conclusion The established PCR-SSP based multiple PCR system is efficient to screen the rare blood group antigens K and Ytb. The information of rare blood donors obtained from the screening can be used to improve the capability of compatible transfusion.