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《Chongqing Medicine》 2017-13
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Ambiguity results distribution and its solutions of HLA-A,B and DRB1 sequence-based typing

Li Hengcong;Pei Yongfeng;Huang Huini;Wu Guoguang;Nanning Blood Center/Nanning Institute of Transfusion Medicine;  
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.
【Fund】: 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)
【CateGory Index】: R440
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