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《Chinese Journal of Blood Transfusion》 2014-01
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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
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【Co-citations】
Chinese Journal Full-text Database 10 Hits
1 ZHOU Haojie;WANG Chuanxi;NIE Yongmei;Guangzhou Blood Center;;Polymorphism of HPA-21 in Han blood donors in Guangzhou,China[J];Chinese Journal of Blood Transfusion;2013-08
2 Yang Lan;Liang Xiuyun;Zeng jianghui;Department of Clinical Laboratory,the Third Affiliated Hospital of Guangxi Medical University/Nanning Municipal Second People's Hospital;;The difference analyses of polymorphism of human platelet antigen 1-17 bw gene system in Yao,Zhuang and Miao nationality of Guangxi[J];International Journal of Laboratory Medicine;2013-17
3 YANG Fang;HUANG Jie;ZENG Xiaojing;YANG Puyu;CONG Shuo;Department of Clinical Laboratory Medicine,Guiyang Medical College;Department of Blood Transfusion,Affiliated Hospital of Guiyang Medical College;;Investigation of the polymorphism of HPA-1~5 gene and the platelet transfusion refractoriness Dong ethnic group in Guizhou province,China[J];Chinese Journal of Blood Transfusion;2013-11
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5 WU Guoguang;ZHOU Yan;ZHONG Zhoulin;LI Lilan;LIU Jinlian;SHEN Weidong;Nanning Institute of Transfusion Medicine;;Experimental study on the platelet transfusion refractoriness mediated by the anti-CD36:four cases reports in China[J];Chinese Journal of Blood Transfusion;2014-01
6 YANG Lan;LIANG Xiu-yun;ZENG Jiang-hui;Department of Laboratory,the Third Hospital of Guangxi Medical University/Nanning Second People's Hospital;;Human platelet antigen gene polymorphism analysis in Yao and Zhuang nationality in Guangxi[J];Laboratory Medicine and Clinic;2013-20
7 MIAO Jing;ZENG Xiao-jing;Department of Hematology,the Affiliated Hospital of Guiyang Medical College;;Correlation between polymorphism of HPA-1~6,15 and the effect of platelet transfusion in patients with acute leukemia[J];Hainan Medical Journal;2014-03
8 TANG Zhao-hui;LI Guo-ying;LIU Yuan-yuan;XUE Shuang-lin;WANG Ming-lu;LI Yuan-rong;Gansu Provincial Blood Center;;Investigation on gene frequency of human platelet antigen-15 system in Gansu[J];Chinese Journal of Health Laboratory Technology;2013-08
9 CHEN Yu-ning;WANG Yan-wei;Department of Laboratory,The G eneral Hospital of Shenyang Military Region;;Genotyping of platelet antigens HPA-2 with melting curve analysis[J];Chinese Journal of Public Health;2013-09
10 LIU Bing-Xian;LIU Yu-Jia;GAO Guang-Ping;WANG Dan;ZHOU Rui-Hua;MA Qiang;ZHANG Yan;YU Xiu-Qing;ZHENG Hua;Blood Bank of Mudanjiang;School of Traditional Chinese Medicine,Capital Medical University;;Applications of Platelet Antigen Gene Typing and Matching in the Platelet Transfusion[J];Journal of Experimental Hematology;2014-01
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