中国人群人类白细胞抗原A、B和DRB1基因测序分型中模棱两可等位基因的鉴别  

Identification of ambiguous allele pairs genotyped by sequencing method for human leukocyte antigens-A, B and DRB1 locus in Chinese population

作　　者：程良红[1] 邹红岩[1] 金士正[1] 李桢[1] 王大明[1] 钱长江[2] 陈甜甜[2] 

机构地区：[1]深圳市血液中心组织配型与免疫遗传研究室,广东省深圳市518035 [2]大连医科大学检验医学系,辽宁省大连市116044

出　　处：《中国组织工程研究与临床康复》2009年第31期6167-6171,共5页Journal of Clinical Rehabilitative Tissue Engineering Research

基　　金：广东省科技计划项目(2008B030301278);广东省医学科研基金项目(A2008622);深圳市科技计划项目(200802117)~~

摘　　要：背景:人类白细胞抗原基因测序分型中,当某些等位基因间的差异碱基位于测序范围外时,无法得到清晰的等位基因结果。目的:分析中国人群人类白细胞抗原A、B、DRB1基因测序分型中模棱两可等位基因的分布规律,探讨其在中华骨髓库大样本人类白细胞抗原A、B、DRB1基因的测序分型中的解决方案。设计、时间及地点:采用随机抽样方法对研究样本进行人类白细胞抗原A、B、DRB1基因的测序分型,并对其中的模棱两可等位基因进行验证性实验,于2006-01/2008-06在深圳市血液中心完成。材料:658名深圳骨髓库汉族供者乙二胺四乙酸盐抗凝血5mL。方法:采用聚合酶链式反应--测序分型方法对658名深圳骨髓库供者的人类白细胞抗原A、B和DRB1基因进行高分辨分型,并采用针对相应位点的高分辨聚合酶链式反应-序列特异性引物方法对其中由于碱基差异位于检测区外而产生的模棱两可等位基因进行鉴别。主要观察指标:模棱两可等位基因的分布及确认。结果:658份标本中,人类白细胞抗原A、B和DRB1三个基因座的模棱两可等位基因分别为9个(2种)、140个(5种)、406个(8种),占等位基因总数的14.06%(555/3948)。高分辨聚合酶链式反应-序列特异性引物鉴定结果显示,中国报道的DRB1*1401被全部确认为DRB1*1454;12例B*0705/B*0706中,有1例被确认为B*0706,其他13种等位基因的鉴定结果分别为A*6801、A*7402、B*2705、B*3501、B*4402、B*5801、DRB1*0101、DRB1*0406、DRB1*0803、DRB1*1101、DRB1*1201、DRB1*1302和DRB1*1502。结论:在中华骨髓库大样本人类白细胞抗原基因的测序分型中可以应用直接鉴别的方法区分模棱两可等位基因,但在临床移植前的高分辨确认试验中则需要采用实验方法进行精确分型。BACKGROUND： Some alleles could not be genotyped clearly due to the different base located outside the sequencing region during the human leukocyte antigens genes genotyped by sequencing method. OBJECTIVE： To analyze the distribution of ambiguous alleles in Chinese population and to investigate the resolution project of ambiguous alleles of the human leukocyte antigens genes （A, B, DRB1） genotyped by sequencing method for large-scale donors in China Marrow Donor Project. DESIGN, TIME AND SETTING： Using random sampling, human leukocyte antigens genes （A, B, DRB1 ） typing was done in samples, and the ambiguous alleles were verified. The experiment was performed at the Shenzhen Blood Center from January 2006 to June 2008. MATERIALS： 5 mL ethylenediaminetetraacetate anticoagulated blood was obtained from 658 donors of Han nationality in the Shenzhen Bone Marrow Bank. METHODS： 658 Chinese Han donors were genotyped by polymerase chain reaction-sequence-based-typing method for their human leukocyte antigens-A, B and DRB1 genes, and then using high-resolution polymerase chain reaction- sequence specific primer method to identify the ambiguous alleles due to their different bases located outside the sequencing region. MAIN OUTCOME MEASURES： Distribution and identification of ambiguous alleles were measured. RESULTS： Among the 658 samples, there were 9 （two types）, 140 （five types） and 406 （eight types） ambiguous alleles for human leukocyte antigens-A, B and DRB1 gene respectively, with 14.06% （555/3 948） in the overall alleles. The high-resolution polymerase chain reaction-sequence specific primer genotyping results showed that all DRB1＊1401 alleles previous reported as dominant allele in China identified as DRB1 ＊1454, and one B＊0706 allele was observed in the 12 ambiguous B＊0705/B＊0706 samples. The other 13 ambiguous allele pairs were A＊6801, A＊7402, B＊2705, B＊3501, B＊4402, B＊5801, DRB1＊0101, DRB1＊0406, DRB1＊0803, DRB1＊1101, DRB1＊1201, DRB1＊1302 and

关 键 词：人类白细胞抗原 聚合酶链式反应-序列特异性引物法 模棱两可 等位基因 

分 类 号：R318[医药卫生—生物医学工程]