荧光标记8个miniSTR及Amelogenin复合扩增体系的建立  

Development of an 8 miniSTR and Amelogenin fluorescent-multiplex system

作　　者：刘超[1] 冯冬亮[2] 刘长晖[1] 

机构地区：[1]广州市刑事科学技术研究所,广东广州510030 [2]中山大学中山医学院法医学系,广东广州510080

出　　处：《中国法医学杂志》2011年第2期99-103,共5页Chinese Journal of Forensic Medicine

基　　金：国家十一五科技支撑计划资助项目(2006BAK07B01)

摘　　要：目的建立非CODIS系统miniSTR以及Amelogenin基因座的荧光复合扩增体系。方法筛选8个多态性高的非CODIS系统miniSTR基因座(D20S1082、D6S474、D12ATA63、D9S1122、D2S1776、D1S1627、D3S4529、D2S441),并结合Amelogenin基因座设计荧光标记引物,优化反应条件,建立复合扩增体系。应用该体系对204份广州地区汉族血样,30个家系样本,及30份降解检材进行检测。结果建立的荧光标记8个miniSTR及Amelogenin复合扩增体系分型结果明确,稳定性好,且所有片段长度均少于200bp,提高了降解检材的分型成功率。在广州汉族人群的累积个人识别率为0.99999993,累积非父排除率为0.992287。结论构建的miniSTR荧光复合扩增体系,操作简便,分型准确,重复性好,对降解检材有效,易于在法医实验室推广应用,可对现有试剂盒起补充作用。Objective To develop a fluorescent multiplex system oi 8 non-CODIS miniSTR and Amelogenln for forensic purpose. Methods Eight highly polymorphic non-CODIS miniSTR loci, D20S1082, D6S474, DI2ATA63, D9S1122, D2S1776, D1S1627, D3S4529 and D2S441 were selected, and a fluorescent multiplex system including these loci and the genderspecific Amelogenin was developed through redesigning primers, labeling fluorescence dye,and optimizing experiment conditions. 204 randomly selected individuals of Guangzhou Han population,members of 30 families and 30 degraded samples were genotyped using this system. Results Tested by the new multiplex system, all the loci can generate stable and full profiles with amplicons less than 200bp in size, which resulted in an increased overall typing success rate for degraded DNA samples. The cumulative power of discrimination and probability of exclusion were 0. 999 999 93 and 0. 992 287 respectively in Guangzhou Han population. Conclusion The established fluorescent nmhiplex system was robust, sensitive and stable, and is of high value in forensic application. It can serve as a new approach for the analysis of degraded DNA as well as an effective supplementary of commercial kits.

关 键 词：法医物证学 MINISTR 复合扩增 降解DNA 

分 类 号：D919[医药卫生—法医学]