3个miniSTR基因座四色荧光复合分型体系的构建及应用  

Construction and application of four fluorescence labeled multiplex typing system for 3 miniSTR loci

作　　者：刘宁[1] 左敏[1] 李淑瑾[1] 丛斌[1] 白雪[1] 谷建立[1] 侯志平[1] 

机构地区：[1]河北医科大学法医学系河北省法医学重点实验室,河北石家庄050017

出　　处：《中国法医学杂志》2009年第3期154-157,共4页Chinese Journal of Forensic Medicine

基　　金：河北省"十一五"科技支撑计划项目(072461479D)

摘　　要：目的建立D3S3053、D6S474、D20S482(扩增片段<150bp)3个miniSTR基因座四色荧光复合分型体系,用于高度降解DNA样本的基因分型。方法采用6-FAM、HEX、TAMRA荧光染料标记D20S482、D3S3053、D6S474基因座上游引物,构建并优化复合扩增体系,在ABI 310遗传分析仪上对扩增产物进行电泳分析,Genemapper 3.2软件分析产物片段大小并进行分型。采用上述体系对120份河北汉族健康无关个体血样进行检测,并计算群体遗传学参数。比较该体系与ID试剂盒用于高度降解检材的成功率及灵敏度。结果D3S3053、D6S474、D20S482 3个miniSTR基因座荧光标记复合扩增分型体系稳定可靠,灵敏度达50pg,用于高度降解检材分析的成功率明显高于ID试剂盒。3个基因座在河北汉族人群中的累积个人识别能力为0.998,累积非父排除率为0.84。结论该系统可用于分析高度降解DNA样本的基因分型,进行法医学个人识别和亲权鉴定。Objective To construct a four fluorescence labeled multiplex typing system for three miniSTR loci D3S3053, D6S474 and D20S482 in which the amplified fragments were shorter than 150bp, and to apply in the analysis of highly degraded DNA samples. Methods The upstream primers of D20S482, D3S3053 and D6S474 were labeled with 6-FAM, HEX, TAMRA respectively, and multiplex PCR system for the three miniSTR loci was established and optimized. The amplified products were detected using ABI 310 Genetic Analyzer. The genotype of each sample was defined according to the length of the amplification products analyzed by GeneMapper Analysis Software 3.2. Samples from 120 unrelated individuals of Hebei Han Population were detected using the established multiplex system to get the population genetic parameters. Highly degraded specimens were also detected using the established multiplex system and ID kit respectively, and the success rate and the sensitivity were compared for the two typing systems. Results The muhiplex-PCR typing system for three miniSTR loci D20S482, D3S3053 and D6S474 showed high stability and sensitivity （50pg）. The success rate for typing highly degraded samples using this system was significantly higher than using ID kit. The cumulative discrimination power of three loci reached to 0.998, and the cumulative chance of exclusion reached to 0.84. Conclusion The multiplex set is useful in personal identification and paternity testing, especially for the degraded DNA samples.

关 键 词：法医物证学 MINISTR 复合扩增 荧光标记 降解检材 

分 类 号：D919[医药卫生—法医学]