Genotyping of human platelet antigens HPA-2,-3 and-15 by multiplex PCR-CE

[Objective] To develop a multiplex PCR-CE method for genotyping of human platelet antigens(HPA)-2,-3 and-15 and investigate gene frequencies of HPA-2,-3 and-15 in Dalian Han population with the method.[Methods] The allele-specific primers and common primers for genotyping of HPA-2,-3 and-15 were designed.The common primers were labeled with fluorescence.The genes of HPA-2,-3 and-15 were amplified using multiplex PCR.Then PCR products were detected using CE.HPAs were genotyped according to different length of PCR products.The multiplex PCR-CE was compared with the conventional PCR-SSP.In addition,dates of HPAs genotyping in Dalian Han population were compared with those in other Chinese populations.[Results] There was complete concordance of results for all samples tested by multiplex PCR-CE and PCR-SSP.The gene frequencies of HPA-2a,-2b,-3a,-3b,-15a and-15b in Dalian Han population were 0.9200,0.0800,0.5750,0.4250,0.5800 and 0.4200 respectively,which did not significantly differ from those in Shan dong,Shanghai and Zhejiang Han populations(P0.05).The gene frequencies of HPA-2,-3 in Dalian Han population did not significantly differ from those in Xinjiang Uighurs population(P0.05),but the gene frequency of HPA-15 in Dalian Han population significantly differed from that in Xinjiang Uighurs population(P0.05).[Conclusions] Multiplex PCR-CE for genotyping of HPAs is a fast,simple and accurate method.There is different in frequency distribution of HPA-15 alleles among different races of China.