人类白细胞抗原-Ⅰ类抗原的DNA分型与临床应用  

DNA typing for HLA A, B antigens by polymerase chain reaction with sequence specific primers and clinical application

作　　者：谭建明[1] 唐孝达[1] 谢桐[1] 徐琴君[1] 丁言德[1] 

机构地区：[1]上海市第一人民医院肾移植中心

出　　处：《中华医学杂志》1998年第10期763-767,共5页National Medical Journal of China

基　　金：卫生部科研基金;上海市医学领先专业重点科研基金

摘　　要：目的采用顺序特异引物聚合酶链反应技术（ＰＣＲＳＳＰ）建立汉族人群白细胞抗原Ⅰ（ＨＬＡⅠ）类抗原Ａ、Ｂ位点ＤＮＡ分型方法。方法设计合成８１个特异性引物和１对阳性对照引物，组成２０个ＰＣＲ反应用于Ａ位点分型、４１个ＰＣＲ反应用于Ｂ位点分型，建立一步法ＰＣＲＳＳＰ方法，应用于６２份标准ＤＮＡ和３４５例肾移植供受者的ＨＬＡⅠ类ＤＮＡ分型。结果所有样本ＰＣＲＳＳＰ基因分型均获得成功，无假阳性和假阴性出现，４０份样本的重复率１００％，总耗时５小时。分型结果经双盲验证完全符合。可准确分辨出Ａ位点等位基因１９个、Ｂ位点等位基因４１个；实际检出汉族人群Ａ抗原特异性１３个、Ｂ抗原特异性３２个。结论ＰＣＲＳＳＰ技术行ＨＬＡⅠ类Ａ、Ｂ位点ＤＮＡ分型，分辨率高、特异性强、重复性好、相对简捷快速，分型结果较血清学方法更加精确可靠。Objective To establish DNA typing for HLA A, B antigens in Chinese by polymerase chain reaction with sequence specific primers (PCR SSP). Methods DNA samples were obtained from 178 unrelated donors and 167 kidney recipients. An additional panel of 62 standard DNAs that were typed by UCLA tissue typing lab in USA. A rapid genotyping for HLA Ⅰ class (A, B antigens) by PCR SSP was set up by designed and synthesized 81 specific primers and 1 pair of internal control primer, combining in 61 one step reactions (20 PCR reactions for A alleles, 41 PCR reactions for B alleles). Results HLA A, B alleles were successfully typed in 345 clinical samples and 62 standard DNAs by PCR SSP technique. No false positive or false negative typing results were obtained. Reproducibility was 100% in 40 samples. The overall time of DNA typing was 5 hours. The typing results were consistent with those of UCLA tissue typing lab. Nineteen alleles of HLA A and 41 HLA B alleles were accurately distinguished. Thirteen HLA A alleles and thirty two HLA B alleles in Chinese were practically typed. Conclusion DNA typing for HLA Ⅰ class (A, B antigens) by PCR SSP has proved to be a technique of high resolution, high specificity, well reproducibility, and more suitable for clinical application than serology.

关 键 词：人白细胞抗原 HLA抗原 DNA分型 PCR 移植免疫学 

分 类 号：R392.4[医药卫生—免疫学] R392.11[医药卫生—基础医学]