Study on multi-PCR-SSP technique for HPA-1 ~ 17bw genotyping and the distribution of HPAs polymorphism in Chinese Zhuang population
SHEN Weidong;LI Lilan;LIAO Yan;ZHAO Tongmao;WU Guoguang;Nanning Institute of Transfusion Medicine;National Institutes of Health,NIH,USA;
Objective To develop a multi-PCR-SSP technique for human platelet antigens(HPAs)-1 to-17bw genotyping.And study on the distribution of HPAs polymorphism in Zhuang population by this technique.Methods We designed various combinations of HPA sequence-specific primers in order to simultaneously amplify several target HPA gene fragments in one PCR reaction by using one multi-primers combination.The results of HPA genotype were read and assigned according to the size of the target fragment and the presence or absence of the fragment.The effectiveness of the multi-PCRSSP technique was evaluated by using 17 HPA reference DNA samples that were provided in 14th Platelet Immunology Workshop and 100 HPA-known DNA samples of platelet donors.Total of 2 659 random samples from unrelated and healthy adults of Zhuang population in Guangxi region were studied for HPA-1 to-17bw genotype and antigens polymorphism by the developed technique.Results There are 16 primers mixes in the multi-PCR-SSP method,and 6 of them are multi-PCR primers combinations mixes in our technical system.100% concordance of genotyping results were showed by the evaluation of using 17 reference DNA samples and 100 DNA samples of known-HPA-genotype platelet donors.The HPAs allele frequencies in Zhuang population were 0.991 5 and 0.008 5 for HPA-1a and-1b;0.956 9 and 0.043 1 for HPA-2a and-2b;0.512 2 and 0.487 8 for HPA-3a and-3b;0.985 0 and 0.015 0 for HPA-5a and-5b;0.985 1 and 0.014 9 for HPA-6a and -6b;0.525 6 and 0.474 4 for HPA-15a and-15b,respectively.There only a / a homozygosis was detected in HPA-4,HPA-7 ~ 14bw,-16 ~ 21bw.And a / a and b / b homozygosis in HPA-2 ~ 3,HPA-6bw and HPA-15 were found.Conclusion In this study,we successfully established a multi-PCR-SSP technique for HPA-1 to-17bw genotyping.It was a rapid and accurate method for HPA genotyping,particularly suitable for large quantity samples' HPA-1 ~ 17bw genotyping.This study showed that the HPAs polymorphism in Zhuang population had their own national characteristics,and the data helps us to know the characteristics of platelet immunological in Zhuang population.
【Fund】： 广西科学研究与技术开发计划项目(主席基金)(081 60-05);; 南宁市科学研究与技术开发计划项目(201003048C-2);; 广西壮族自治区卫生厅自筹经费科研课题(Z2010197)
【CateGory Index】： R394
【CateGory Index】： R394