16个Y-STR基因座快速复合扩增体系的构建研究  

Study on Construction of a 16-Plex Rapid Y-STR Amplification System

作　　者：董倩[1] 欧元[2] 韩俊萍 李彩霞[2] 尚蕾[2] 赵蕾[2] 丁光树 孙辉[2] 李万水[2] 叶健[2] 朱波峰[1] 孙敬[2] DONG Qian;OU Yuan;HAN Jun-ping;LI Cai-xia;SHANG Lei;ZHAO lei;DING Guang-shu;SUN Hui;LI Wan-shui;YE Jian;ZHU Bo-feng;SUN Jing(Key Laboratory of Shaauxi Province for Craniofacial Precision Medicine Research;Clinical Research Center of Shaauxi Province for Dental and Maxillofacial Diseases, College of Stomatology, Xi＇an Jiaotong University, Xi＇an 710004, Shaauxi, Chin;2. Beijing Engineering Research Center of Crime Scene Evidence Examinatio;Key Laboratory of Forensic Genetics, Institute of Forensic Science, Ministry of Public Security, Beijing 100038, Chin;3. Technology Department of Chaoyang Sub- bureau, Beijing Public Security Bureau, Beijing 100025, China)

机构地区：[1]西安交通大学口腔医学院,陕西省颅颌面精准医学研究重点实验室,陕西省牙颌疾病临床医学研究中心,中国陕西西安710004 [2]公安部物证鉴定中心,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,中国北京100038 [3]北京市公安局朝阳分局刑事侦查支队,中国北京100025

出　　处：《生命科学研究》2018年第2期99-113,共15页Life Science Research

基　　金：“十三五”国家重点研发计划项目(2017YFC0803507);公安部技术研究计划项目(2016JSYJC11)

摘　　要：为了构建一套16个Y-STR基因座的快速复合扩增体系,用于满足公安的实战时效性需求,文中选择DYS19、DYS385a/b、DYS390、DYS391、DYS392、DYS393、DYS438、DYS439、DYS437、DYS448、DYS456、DYS458、DYS635、Y_GATA H4和DYS447共16个基因座,将Roche FastStart Taq DNA Polymerase体系与ProPlex热循环仪结合,以法医DNA标准品9948为实验模板,从扩增条件、扩增体积、缓冲液选择、DNA聚合酶用量、基因座间的平衡性调整、快速扩增程序的优化等方面进行一系列复合扩增实验,比较不同条件下等位基因的丢失、峰高、基因座间峰的均衡性及非特异峰。结果发现,该快速体系30 min即可获得样本全部16个Y-STR基因座分型,且各等位基因间均衡性良好,无非特异性峰。以上信息初步表明建立的16个Y-STR基因座的快速复合扩增体系可明显提高样品检测效率,对实战中一些时效性案件具有一定的实际意义。To establish a 16-plex rapid Y-STR amplification system for the actual needs of public security, sixteen STR loci, including DYS19, DYS385a/b, DYS390, DYS391, DYS392, DYS393, DYS438, DYS439, DYS437, DYS448, DYS456, DYS458, DYS635, Y_GATA H4 and DYS447, were chosen based on the cur- rent Y-STR genotyping for development of the rapid PCR. Roche FastStart Taq DNA Polymerase and DNA standard sample 9948 were used during an amplification and optimization process. The 16-plex rapid Y- STR amplification system was achieved by performing various experiments, including choosing amplification conditions and the volume of DNA polymerase, and adjusting interlocus balance, thermal cycling parameters and the volume of rapid PCR system, reaction buffers and a lot of additives. The results showed that, through this 16-plex rapid Y-STR amplification system, all 16 samples of Y-STR loci could be obtained in 30 min- utes, with a good balance between the alleles and without nonspecific amplification. It proved that the rapid recombination amplification system of 16 Y-STR sites can improve the efficiency of sample detection and is of practical significance for the timeliness of special cases in public security.

关 键 词：法医DNA 快速PCR 快速Y-STR复合扩增 16个基因座 遗传多态性 

分 类 号：Q3-3[生物学—遗传学]