摘要
目的实现以荧光标记复合扩增产物为扩增模板的STR序列多态分型。方法筛选常用STR分型试剂盒基因座并设计引物,构建常染色体STR复合扩增体系。分别以毛细管电泳(CE)试剂盒GlobalFilerTM、PowerPlex?21和IdentifilerTM Plus的扩增产物为模板,扩增、建库测序及数据分析,评估体系分型准确性及对CE扩增产物的通用性,并与Precision ID GlobalFilerTM NGS STR Panel体系进行比较。结果该复合扩增体系能够从3款CE试剂盒的扩增产物中获得所有基因座的正确分型和序列多态信息,其体系均衡性和杂合子均衡性亦良好,且均优于Precision ID体系。结论该复合扩增体系建立了CE技术与二代测序技术之间的桥梁,实现了检材的零利用,能够进一步挖掘荧光标记扩增产物的STR序列多态信息,是对现有技术的提升和有效补充。
Objective To establish a 13 autosomal STR multiplex system for detecting the sequence-based STR genotype of CE PCR products.Methods PCR products of CE kit GlobalFilerTM,PowerPlex?21,IdentifilerTM Plus were used as the template to estimate the precision and universality of the 13 autosomal STR multiplex system after it was established.And its performance was compared to the Precision ID GlobalFilerTM NGS STR Panel.Results All the loci,both in the 13 autosomal STR multiplex system and the CE kit,with their concordant length-based genotype and sequence were obtained.It got credible results from the 13 autosomal STR multiplex system for its good performance of the loci coverage ratio and the allele coverage ratio.Conclusion The 13 autosomal STR multiplex system built a bridge between CE technology and NGS technology,it could not only use the CE products but also explore the STR sequence polymorphic information of CE products,which elevated and supplemented the genotype technology effectively.
作者
郭立亮
吴浩
刘宗伟
龚辰宇
张驰
康克莱
韦美甜
孙宏钰
季安全
王乐
Guo Liliang;Wu Hao;Liu Zongwei;Gong Chenyu;Zhang Chi;Kang Kelai;Wei Meitian;Sun Hongyu;Ji Anquan;Wang Le(Department of Forensic Medicine,Zhongshan School of Medicine,Sun Yat-sen University,Guangzhou 510080,Guangdong,China;National Engineering Laboratory for Forensic Science&MPS Key Laboratory of Forensic Genetics&Institute of Forensic Science,Ministry of Public Security'(MPS),Beijing 100038,China;Changshu Public Security Bureau,Changshu 215500,Jiangsu,China;School of Forensic Medicine,Kunming Medical University,Kunming 650500,China)
出处
《中国法医学杂志》
CSCD
2021年第3期263-268,共6页
Chinese Journal of Forensic Medicine
基金
公安部技术研究计划(2019JSYJA05)
公安部科技强警基础工作专项(2019GABJC15)
公安部物证鉴定中心基本科研业务费(2019JB009,2019JB045)。
关键词
法医遗传学
二代测序
短串联重复序列
序列多态性
毛细管电泳扩增产物
Forensic genetics
Next generation sequencing(NGS)
Short tandem repeat(STR)
Sequence polymorphism
CE PCR products
