摘要
目的调查广西人群HLA-A、B、DRB1基因测序分型中模棱两可结果的分布状况,提出解决方案。方法采用聚合酶链反应-直接测序分型方法对1 000名中华骨髓库广西分库供者的HLA-A、B、DRB1基因进行测序分型,分析3个基因座模棱两可分型结果的分布情况,并分别采用高分辨聚合酶链反应-序列特异性引物法和组特异性测序引物法进行解决。结果 1 000份标本中,96.7%的标本HLA-A、B、DRB1基因至少有1个位点出现模棱两可结果,其中HLA-A、B、DRB1各位点出现模棱两可结果的比例分别为65.7%、58.8%、77.2%。对于检出的模棱两可结果标本,单独采用组特异性测序引物法可以解决87.37%HLA-A、93.54%HLA-B和60.49%的HLA-DRB1基因模棱两可分型结果,使用高分辨聚合酶链反应-序列特异性引物法解决12.63%HLA-A、4.76%HLA-B和15.29%HLA-DRB1基因模棱两可分型结果,剩余1.70%HLA-B和24.22%的HLA-DRB1基因模棱两可分型结果联合使用组特异性测序引物法和高分辨聚合酶链反应-序列特异性引物法解决。结论组特异性测序引物法和高分辨聚合酶链反应-序列特异性引物法分别对位于HLA-A、B和HLA-DRB1基因检测区内、外的模棱两可分型结果具有较高的解决能力,二者互为补充,可有效解决HLA高分辨基因分型结果模棱两可问题。
Objective To investigate the ambiguity results distribution of HLA-A,B and DRB1 gene sequence-base typing in Guangxi population and to propose the way to resolve.Methods HLA-A,B and DRB1 genes of 1 000 donors in the Guangxi branch bank of China′bone marrow bank were genotyped by PCR-SBT,and then the ambiguity results distribution of the three loci was analyzed.The typing ambiguities results were resolved by high-resolution polymerase chain reaction-sequence-specific primers（PCRSSP）and group specific sequencing primer（GSSP）methods,respectively.Results Among 1 000 samples,at least 1locus in HLAA,B and DRB1 genes in 96.7% samples appeared the ambiguity results,in which the proportions of HLA-A,B and DRB1 loci appearing ambiguity results were 65.7%,58.8% and 77.2% respectively.For the samples of detected ambiguity results,single using the GSSP method could resolve the ambiguity typing results of 87.37% HLA-A,93.54% HLA-B and 60.49% HLA-DRB1,using high-resolution PCR-SSP could resolve the ambiguity typing results of 12.63% HLA-A,4.76% HLA-B and 15.29%HLA-DRB1,and the rest 1.70%HLA-B and 24.22%HLA-DRB1 ambiguity results were resolved by both GSSP and high-resolution PCR-SSPs method.Conclusion GSSP and high-resolution PCR-SSPs methods have high abilities to solve HLA ambiguity results both locate inside and outside the sequencing region,respectively.GSSP and high-resolution PCR-SSPs methods are supplement for each other,which can effectively resolve the problem of ambiguity results in high resolution HLA typing.
作者
李恒聪
裴永峰
黄惠妮
吴国光
Li Hengcong Pei Yongfeng Huang Huini Wu Guoguang(Nanning Blood Center /Nanning Institute of Transfusion Medicine, Nanning,Guangxi 530007 ,Chin)
出处
《重庆医学》
CAS
北大核心
2017年第13期1759-1761,共3页
Chongqing medicine
基金
广西壮族自治区南宁市科学研究与技术开发计划项目(201003048C-3)
广西壮族自治区卫生厅科技研究计划项目(Z2010198)
