基于二代测序技术分析多拷贝DYF404S1基因座遗传多态性初步研究  

A preliminary study on genetic polymorphisms of multi-copy DYF404S1 locus based on the next generation sequencing

作　　者：安雷雷 郭利伟 钱嘉林[3] 于春蕊 赵晶[4] 严江伟[4] 焦会永 黄艳梅 An Leilei;Guo Liwei;Qian Jialin;Yu Chunrui;Zhao Jing;Yan Jiangwei;Jiao Huiyong;Huang Yanmei(Department of Forensic Genetics,School of Forensic Medicine,Xinxiang Medical University,Xinxiang 453003,China;Department of Anatomy of Sanquan College,Xinxiang Medical University,Xinxiang 453003,China;Beijing Center for Physical ＆ Chemical Analysis,Beijing 100094,China;Beijing Institute of Genomics Chinese Academy of Sciences,Beijing,100029,China;Xinxiang Key Laboratory of Forensic Science Evidence,School of Forensic Medicine,Xinxiang Medical University,Xinxiang 453003,China)

机构地区：[1]新乡医学院法医学院法医物证学教研室,河南新乡453003 [2]新乡医学院三全学院人体解剖学教研室,河南新乡453003 [3]北京市理化分析测试中心,北京100094 [4]中国科学院北京基因组研究所,北京100029 [5]新乡医学院法医学院新乡市法庭科学证据重点实验室,河南新乡453003

出　　处：《中国法医学杂志》2018年第5期485-488,484,共5页Chinese Journal of Forensic Medicine

基　　金：河南省教育厅自然科学基金资助项目13A310849;河南省教育科学"十二.五"规划立项重点课题2013-JKGHB-0035;新乡医学院培育基金重点项目2013ZD114

摘　　要：目的基于二代测序技术(next generation sequencing, NGS)用于Y染色体多拷贝短串联重复序列(multi-copy short tandem repeat on Y chromosome, Multi-copy Y-STRs)DYF404S1分型方法建立及在河南汉族群体中遗传多态性分析的初步研究。方法在DYF404S1基因座特异引物两端添加适宜的接头,得到带有不同接头的引物组合,共25组。采用BIOO RAPID DNA试剂盒构建二代测序文库,使用Illumina MiseqTM二代测序仪对225例河南汉族群体的男性样本进行测序,STRait Razor V2.0软件进行测序数据分析,同时与一代测序ABI3100遗传分析仪分型结果比对分析。结果基于Illumina MiseqTM二代测序平台,采用引物两端加接头的方法对225例河南汉族无关男性个体样本进行DYF404S1基因座分型,其中220例成功分型,成功率达97.8%;同时使用3130遗传分析仪进行DYF404S1基因座分型,分型结果与二代测序分型结果一致。3例样本出现等位基因异常分型,均表现为3等位基因模式。DYF404S1基因座在河南汉族群体中的单体型多样性(haplotype diversity, HD)和个体识别力(discrimination power, DP)分别为0.9356和0.9311。结论初步建立了NGS技术用于多拷贝DYF404S1分型方法,为法医学实践和群体遗传学提供了技术和理论基础。Objective Establishment of a method for genotyping multi-copy STR DYF404S1 locus on Y chromosome based on the next generation sequencing （NGS） and researches on its genetic polymorphism in Henan Han population were conducted. Methods The self-designed adapters were added on the both ends of specific primer of DYF404S1 locus, Primers were divided into 25 groups totally. The library was constructed by using BIOO RAPID DNA genetic library kit. The sequence of alleles of DYF404S 1 locus were carried out by Illumina MiseqTM sequencing kit. Sequence data were analyzed by STRait Razor V2.0 software. The concordance rate was measured between Illumina MiseqTM kit and CE-based STR profiling kits. Results Among 225 unrelated males blood DNA samples from Henan Han population, 220 samples were typed successfully based on NGS platform, the successful rate was 97.8%. The genotyping of DYF404S1 showed no difference between NGS and CE based assay. AUelic Abnormal Typing appeared in 3 Samples,all of which showed tri-allelic patterns. The haplotype locus in the Han population of Henan were 0.935 and DYF404S 1 locus genotyping by NGS was established, forensic practice and population genetics. diversity and the discrimination power of multi-copy DYF404S1 0.931, respectively. Conclusions The method for Multi-copy which could provide a technical support and theoretical basis for

关 键 词：法医物证学 多拷贝 DYF404S1基因座 Y染色体 二代测序技术 

分 类 号：D919[医药卫生—法医学]