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目的 研究建立快速等位基因特异性 (AS)引物 PCR技术 ,同时对人血小板同种异型抗原系统 (HPA 1,2 ,3,4,5 )等位基因进行分型。方法 设计合成 15条等位基因特异性引物 ,摸索最适引物浓度、Mg+ + 浓度及扩增参数。用该技术对 10 0名北京地区健康献血者HPA 1～ 5系统等位基因进行分型。结果 从 10 0名北京地区献血员观察到的HPA基因频率分别是HPA 1a和 1b为 0 .995和 0 .0 0 5 ,HPA 2a和 2b为 0 .90 0和 0 .10 0 ,HPA 3a和 3b为 0 .775和 0 .2 2 5 ,HPA 4a和 4b为 1.0 0 0和 0 ,HPA 5a和 5b为 0 .970和 0 .0 30。结论 该方法简单、快速 ,结果准确 ,适用于临床及常规献血员血小板分型。
OBJECTIVE: To establish a rapid allele-specific (AS) primer PCR technique and to classify the human platelet alloantigen system (HPA 1,2, 3,4,5) alleles. Methods Fifteen allele-specific primers were designed and synthesized, and the optimal primer concentration, Mg + concentration and amplification parameters were determined. The technique was used to genotype the HPA 1-5 alleles of 10 healthy blood donors in Beijing. Results The frequencies of HPA genes observed from blood donors in Beijing of 10 0 were 0 .995 and 0 0 5 for HPA 1a and 1b, 0 .90 0 and 0 .10 0 for HPA 2a and 2b respectively, HPA 3a and 3b are 0.775 and 0.225, HPA 4a and 4b are 1.000 and 0, and HPA 5a and 5b are 0.970 and 0.030, respectively. Conclusion The method is simple, rapid, accurate and suitable for clinical and routine blood donors platelet typing.