基于NGS方法构建人类STR、SNP联合检测体系及其性能研究  

Construction and validation a STR/SNP multiplex detection system based on next generation sequencing

作　　者：聂昊 申君毅 郑立 郭茂平 李梦 王蓉 吴轩 陈浩东 张凯凯 李琛 胡欢 郭晋荣 杨瑞琴[1] Nie Hao;Shen Junyi;Zheng Li;Guo Maoping;Li Meng;Wang Rong;Wu Xuan;Chen Haodong;Zhang Kaikai;Li Chen;Hu Huan;Guo Jinrong;Yang Ruiqin(Chinese People's Security University,Key Laboratory of Forensic Genetics,Institute of Forensic Science,Ministry of Public Security,Beijing 100038;Institute of Forensic Science,Public Security Department of Shanxi,Jinzhong,Shanxi 030600,P.R.China;Beijing Microread Genetics Co.,Ltd.,Beijing 100083;Shanxi Police College,Taiyuan,Shanxi 030401,P.R.China)

机构地区：[1]中国人民公安大学,公安部鉴定中心,北京100038 [2]山西省公安厅科技处,山西晋中030600 [3]北京阅微基因技术股份有限公司,北京100083 [4]山西警察学院,山西太原030401

出　　处：《中国法医学杂志》2023年第2期161-165,共5页Chinese Journal of Forensic Medicine

基　　金：中央级公益性科研院所基本科研业务费“新型连锁遗传标记的开发验证与法医物证应用研究”(2022JB019);刑事技术“双十计划”重点攻关任务(2020SSGG0404);大麻地域溯源体系构建及特异性分子标记数据库建设(2021JB002)。

摘　　要：目的 使用NGS方法构建人类STR和SNP遗传标记联合检测体系,并对体系性能做出评价。方法 通过两轮PCR方法构建了同时检测78个STR和233个SNP的测序文库,通过覆盖度和均衡性评价了panel体系,并且开发新的侧翼检测和分型方法从灵敏度、准确性、适用PCR机型、种属特异性、检材类型、模拟降解检材、混合样本区分、批量样本的表现等方面对体系的效能进行了研究。结果 该体系表现较高覆盖度和准确性。新的分析方法,使得在真实检材样本中和CE的一致率超过99.6%。本体系适用广泛的PCR机型和多种检材类型;SNP灵敏度可达到0.1 ng,STR在投入量为0.5 ng时,检出率依然可以达到93.3%;非人物种的干扰几乎为0;能够有效的区分低频率的混合样本(低于1:19)。结论 上述结果表明该体系具有较好的性能,可用于法庭科学实际案件或数据库建设的检测。Objective To establish a combined detection system for human STR and SNP genetic markers based on the next generation sequencing(NGS) method,and to evaluate the efficiency characteristics of the system.Methods In this system,sequencing libraries were constructed by two-step PCR to simultaneously detect 78 STRs and 233 SNPs.The panel was evaluated by analyzing the coverage and depth balance of the sequencing results.We explored the efficiency characteristics of the system,including sensitivity,accuracy,applicability for different PCR instruments,species specificity,specimen type,simulated degradation specimen,mixed sample differentiation and performance of batch samples.Results This system has a high coverage rate and accuracy.The consistency rate between the established detection system and the classical CE(capillary electrophoresis) detection system was more than 99.6 % when practical samples were analyzed.It was applicable to a wide range of PCR instruments and a variety of sample types.The sensitivity for SNP was 0.1ng,and the detection rate of STR was 93.3 % when the amount of the STR sample was 0.5ng.The possibility of the interference of non-human species was almost zero.It could effectively distinguish the mixed samples with low frequency(less than 1:19).Conclusion The above results indicate that the system has good performance and can be used for the detection in practical cases or for database construction.

关 键 词：法医遗传 高通量测序NGS STR SNP 效能验证 法医学应用 

分 类 号：D919.2[医药卫生—法医学]