五色荧光标记20个基因座复合扩增体系及法医学应用

摘要

目的 建立20个基因座五色荧光标记复合扩增检测体系,并评价其法医学应用价值.方法 收集368份无关人血样及55份实际案例样本(包括血斑、体液斑、组织及毛发),采用五色荧光素标记技术,对Amelogenin和19个STR基因座(D19S433、D5S818、D21S11、D18S51、D6S1043、D3S1358、D13S317、D7S820、D16S539、CSF1PO、Penta D、vWA、D8S1179、TPOX、Penta E、TH01、D12S391、D2S1338和FGA)进行基因型检测,并考察方法的一致性、灵敏度、种属特异性及检材适用性.结果 本文五色荧光标记复合扩增检测体系可对所选20个基因座分型,结果稳定准确,且均衡性良好、无杂峰；群体调查显示累积个人识别率和累积非父排除率分别是0.999999999999999999999和0.99999999；灵敏度达125pg,种属特异性高,实际案例检材分型成功率高.结论 本文五色荧光标记复合扩增检测体系各项指标可达到当前商品化试剂盒的检测水平,具有重要的法医学应用价值.%Objective To construct a 20-locus STR fluorescent-multiplex PCR for forensic purposes. Methods 368 unrelated individual blood samples and 55 routine case samples (including bloodstains, body fluid stains, tissues, and hairs) were collected. Amelogenin and 19 STR loci (D19S433, D5S818, D21S11, D18S51, D6S1043, D3S1358, D13S317, D7S820, D16S539, CSF1PO, PentaD, vWA, D8S1179, TPOX, Penta E, TH01, D12S391, D2S1338 and FGA) were used to construct a 20-plex PCR system using a fivecolor fluorescence labeling technique, and then the consistency, sensitivity, species-specificity and biological samples applicability were evaluated. Results The 20-locus STR fluorescent-multiplex PCR system was of well accuracy, stability and balance, which showed rich genetic information ( the cumulative discrimination power and cumulative chance of exclusion reached to 0. 999 999 999 999 999 999 999 and 0. 999 999 99 respectively) , high sensitivity (125pg), high species-specificity and high genotyping success rate for routine biological samples. Conclusion The performance of this 20-locus STR fluorescent-multiplex PCR system can win that of the current commercial kits, and it has important application value in forensic science.