二代测序与毛细管电泳技术STR分型差异现象探究  

Exploring into Inconsistency between STR Genotypes Tested from Next Generation Sequencing and Capillary Electrophoresis

作　　者：郭江玲 尚蕾[2] 李万水[2] 丁光树 杨帆[2] 孙敬[2] 孙辉[2] 张更谦[1] GUO Jiangling;SHANG Lei;LI Wanshui;DING Guangshu;YANG Fan;SUN Jing;SUN Hui;ZHANG Gengqian(Shanxi Medical University,Taiyuan 030001,China;Institute of Forensic Science,Ministry of Public Security(MPS)&National Engineering Laboratory for Forensic Science&Beijing Engineering Research Center of Crime Scene Evidence Examination&MPS’Key Laboratory of Forensic Genetics,Beijing 100038,China)

机构地区：[1]山西医科大学,太原030001 [2]公安部物证鉴定中心,现场物证溯源技术国家工程实验室,北京市现场物证检验工程技术研究中心,法医遗传学公安部重点实验室,北京100038

出　　处：《刑事技术》2022年第2期156-161,共6页Forensic Science and Technology

基　　金：中央级公益性科研院所基本科研业务费专项资金项目(2019JB033、2020JB006);公安部科技强警基础工作专项(2019GABJC16)。

摘　　要：二代测序相较于毛细管电泳技术(capillary electrophoresis, CE),以其体系中可容纳更多的基因座而在法医学实践中更具潜力和价值。MiSeq FGx^(TM)系统是专为法医学设计研发的一个测序平台,其配套试剂盒Forenseq DNA Signature Prep kit具有较高的灵敏度和准确度。本实验利用该试剂盒对41份家系样本进行测序,结果发现,58个STR基因座中有26个基因座出现了等位基因亚型,等位基因总数目增加了79个;与CE一致性上,有1份样本在DYS392基因座出现缺失,推测应与试剂盒引物扩增效率有关,另外还有8份样本在DXS7132基因座出现与CE不一致的情况,通过Sanger测序以及使用其他软件分析后,排除扩增失败,确认是生物信息分析问题。实验证明,二代测序相比于毛细管电泳技术有很多优势,不过目前在数据分析方面还存在一些问题,需要不断优化完善所涉及的生物信息分析方法。随着未来相关技术与标准的不断完善,二代测序会逐渐应用于法医学实践中。Compared with capillary electrophoresis(CE) technology, next generation sequencing(NGS) can accommodate more STR genetic loci within the relevant system, therefore emerging as a promising and potential technology for forensic practice. Illumina’s MiSeq FGxTM system is a sequencing platform dedicated to forensic services, with its supporting kit-Forenseq DNA Signature Prep kit having achieved higher sensitivity and accuracy. 41 pedigree samples were here reported of being sequenced with the kit mentioned above. The results showed that 26 of the tested 58 STR loci had come into being allelic subtypes, bringing forth the total number of alleles to increase by 79. There was one sample showing a dropout at the DYS392 locus against to its CE presentation, being speculated of correlativity with amplifi cation effi ciency of the kit’s primers. Besides, eight samples were found of inconsistency with CE results at the DXS7132 locus. Sanger sequencing was thus conducted and data were analyzed with other software to explore the causation. It was confi rmed that the inconsistency resulted from irrelevant bioinformatics analysis. This study has proved that NGS owns many advantages over CE although it still awaits solutions about some issues in bioinformatics analysis. With the ever-coming continuous improvement of related technologies and standards, NGS is believed to be gradually applied into forensic practice.

关 键 词：法医遗传学 二代测序技术 毛细管电泳 一致性 DXS7132 生物信息分析 

分 类 号：DF795.2[医药卫生—法医学]