快速AS-PCR技术用于血小板HPA-1,-2,-3,-4,-5抗原系统等位基因的分型  

Rapid determination of platelet alloantigen genotypes of HPA-1, 2, 3, 4, 5 systems by AS-PCR method

作　　者：单小燕[1] 张志欣[1] 李伟[1] 陈素媛[1] 志宏[1] 袁庆珍 

机构地区：[1]北京市红十字血液中心HLA实验室

出　　处：《中华微生物学和免疫学杂志》2000年第4期377-380,共4页Chinese Journal of Microbiology and Immunology

摘　　要：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 develope a rapid allele-specific PCR(AS-PCR) method for simultaneous genotyping of human platelet antigens(HPA) 1,2,3,4,5 systems. Method Fifteen allele-specific primers were synthesized and used in the AS-PCR typing of HPA-1, 2, 3, 4, 5 from 100 randomly selected donors in Beijing. Result The HPA gene frequencies for HPA-1a and 1b, PHA-2a and 2b, PHA-3a and 3b, HPA-4a and 4b, HPA-5a and 5b, respectively were 0.995 and 0.005, 0.900 and 0.100, 0.775 and 0.225, 1.000 and 0, 0.970 and 0.030. Conclusion The AS-PCR method is simple, rapid and accurate, can therefore be used in clinical HPA genotyping.

关 键 词：等位基因特异性 聚合酶链反应 血小板同种抗原 

分 类 号：R392[医药卫生—免疫学]