应用下一代测序技术进行HLA-A、HLA-B基因分型的模棱两可结果情况分析  

Analyze the ambiguous results of HLA-A, -B genotyping using next generation sequencing method

作　　者：董丽娜[1,2] 赵烁贤 王芳 章伟[1,2] 何吉 朱发明[1,2] DONG Li-na;ZHAO Shuo-xian;WANG Fang;ZHANG Wei;HE Ji;ZHU Fa-ming(Blood Center of Zhejiang Province,Hangzhou,Zhejiang 310052,China)

机构地区：[1]浙江省血液中心,浙江杭州310052 [2]浙江省血液安全研究重点实验室,浙江杭州310052

出　　处：《中国卫生检验杂志》2019年第16期1924-1927,1931,共5页Chinese Journal of Health Laboratory Technology

基　　金：浙江省科技厅公益技术应用研究(2017C33121);浙江省医药卫生平台重点资助计划(2015ZDA012)

摘　　要：目的统计分析NGS法进行HLA-A、HLA-B基因分型检测的模棱两可结果的种类、比例。方法对369例浙江省脐带血库标本采用NGS法检测HLA-A、HLA-B位点,并同步对其中94例标本采用PCR-SBT测序法测定HLA-A、HLA-B位点,余下275例标本采用PCR-SSO流式磁珠法检测。使用专业软件指定NGS法的HLA-A、HLA-B分型结果,直接计算法分析模棱两可组合比例。结果所有标本NGS法检测结果与PCR-SBT法或PCR-SSO法相符合,未发现NGS法遗漏等位基因情况。NGS法以HLA-A、HLA-B命名*后第四区域的数字作为高分辨水平统计,369例有34例标本出现模棱两可结果,占9.21%(34/369),其中只有2例标本HLA-A、HLA-B位点同时存在模棱两可结果。HLA-A位点有9例标本存在模棱两可结果,表现出8种类型;HLA-B位点有27例,25种类型,分别占标本总数的2.44%(9/369)、7.32%(27/369)。结论应用NGS技术仍有少量HLA-A、HLA-B基因分型的模棱两可组合结果。Objective To statistically analyze the characteristics of ambiguous results of the HLA-A,HLA-B genotyping using next generation sequencing(NGS)method.Methods 369 samples from Zhejiang Cord Blood Bank were genotyped for HLA-A,HLA-B loci by NGS method,and meanwhile the HLA-A,HLA-B loci of 94 samples among them were detected by PCR-SBT method,and other 275 samples were detected by PCR-SSO flow fluorescent bead method.The genotyping results of HLA-A,HLA-B loci using NGS method were assigned by professional analytical software.The ambiguous combination ratio of the results was calculated by the direct count method.Results The results of all samples using NGS method were consistent with those of PCR-SBT method or PCR-SSO method,and no alleles dropout were found using NGS method.Ambiguous results were found in 34 of 369 samples,accounting for 9.21%(34/369)when HLA-A,HLA-B loci nomination was reach to the fourth field using NGS method for genotyping,including only 2 samples both in HLA-A and HLA-B loci with ambiguous results.There were 9 samples in HLA-A loci with ambiguous results and 8 various types,27 samples in HLA-B loci and 25 types.They accounted for 2.44%(9/369),7.32%(27/369)of all tested samples respectively.Conclusion There are still a few HLA-A,HLA-B genotyping ambiguous combinations results using NGS technology.

关 键 词：下一代测序法 HLA-Ⅰ类基因 模棱两可 测序分析 

分 类 号：R450[医药卫生—治疗学]