多重聚合酶链反应用于人血小板抗原-2、-4、-5系统基因分型的研究

Study on the genotyping of human platelet antigen-2,-4,-5 system by multiplex polymerase chain reaction

目的建立用于检测人血小板抗原(HPA)-2、-4、-5系统基因型的多重聚合酶链反应(PCR)。方法在一个反应体系中同时扩增HPA-2、-4、-5系统特异性目的基因片段。用琼脂糖凝胶电泳确定HPA-2、-4、-5系统基因型;再用多重PCR对75名健康的单采血小板者进行HPA-2、-4-、5系统基因分型,分型结果与PCR-序列特异性引物(PCR-SSP)获得的结果进行比较。结果HPA-2、-4、-5系统分型的结果为:70名为2 a/2 a型,5名为2 a/2b型;73名为4 a/4 a型,2名为4 a/4b型;66名为5 a/5 a型,9名为5 a/5b型。未发现2b/2b、4b/4b及5b/5b纯合子个体。多重PCR结果与PCR-SSP方法获得的结果一致。结论多重PCR具有操作简便、快速、准确等特点,可以用于血小板血型抗原基因分型。

Objective To develop a method for genotyping of the human platelet antigen （HPA） -2,-4,-5 system by means of multiplex polymerase chain reaction （ PCR ） technique . Methods HPA-2,-4,-5 antigen system genes were simultaneously amplified in one amplification system. The results of genotyping HPA-2,-4,-5 antigen system by multiplex PCR were compared with those obtained by PCR-sequence specific primer（SSP）. Results The genotype of HPA-2, -4 ,-5 antigen system obtained by multiplex PCR in 75 healthy platelet donors were 70 of 2a/2a,5 of 2a/2b ,0 of 2b/2b; 73 of 4a/4a,2 of 4a/4b, 0 of 4b/4b; 66 of 5a/5a,9 of 5a/5b and 0 of 5b/5b. All were in agreement with those determined by PCR-SSP. Conclusions The method of multiplex PCR is convenient, economical, rapid,and accurate. It may be appropriate for genotyping of HPA.