用顺序特异引物聚合酶链反应技术行HLA-A抗原DNA分型  

DNA Typing for HLAA by Polymerase Chain Reaction with SequenceSpecific Primers

作　　者：谭建明[1,2] 唐孝达[1,2] 

机构地区：[1]上海市第一人民医院肾移植中心 [2]华西医科大学第一附属医院泌尿外科

出　　处：《华西医科大学学报》1999年第2期133-137,共5页Journal of West China University of Medical Sciences

基　　金：中国博士后科研基金;卫生部科研基金

摘　　要：作者采用顺序特异引物聚合酶链反应技术（ＰＣＲ－ＳＳＰ），建立了汉族人群ＨＬＡ－Ａ位点ＤＮＡ分型方法。研究采用ＰＣＲ－ＳＳＰ对３４５例肾移植供受者样本ＤＮＡ行ＨＬＡ－Ａ位点基因分型均获得成功。共检出Ａ位点等位基因总数６３５个，其中纯合子５５例，杂合子２９０例；可准确分辨ＨＬＡ－Ａ位点等位基因２０个，无假阳性和假阴性出现，重复率１００％，总耗时５小时。分型结果经标准ＤＮＡ验证和美国加州大学配型中心双盲验证完全符合。与标准血清学方法比较显示，６９份血清学空白位点中１６份存在第二个位点，１３份血清学分型错误，血清学总误差率９．０％。由此表明，用该研究建立的ＰＣＲ－ＳＳＰ方法行ＨＬＡ－Ａ位点ＤＮＡ分型，具有高分辨度、高特异性和简捷快速的特点，分型结果较血清学方法更加精确可靠，适合于临床应用。The authors have established a method of DNA typing for HLAA antigen in Chinese by using polymerase chain reaction with sequencespecific primers(PCRSSP).HLAA alleles were successfully typed in all clinical samples and 36 standard DNAs by PCRSSP.A total of 635 A antigens were typed in 345 donorrecipients, including 55 homozygotes and 290 heterozygotes. Twenty alleles of HLAA were accurately distinguished by DNA typing. No false positive or false negative typing results were obtained. Reproducibility was 100%. The overall time of DNA typing was 5 hours. The typing results were concordant with the results of UCLA Tissue Typing Lab. Comparative study of HLAA typing was carried out using PCRSSP and standard serology. The results showed that the discrepancy rate of serology was 9.0%, containing 13 individuals incorrectly interpreted by serology and 16 serological blanks tumed out to be a definable alleles by PCRSSP.In conclusion, DNA typing for HLAA by PCRSSP has proved to be a highresolution,highspecific

关 键 词：聚合酶链反应 器官移植 HLA-A抗原 DNA分型 

分 类 号：R617[医药卫生—外科学] R446.9[医药卫生—临床医学]