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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0.999 999 999 999 999 999 999和0.999 999 99;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 establish a multiplex amplification system for detecting 20 loci and to evaluate its value in forensic medicine. Methods A total of 368 unrelated human blood samples and 55 actual samples (including blood spots, body fluid spots, tissues and hair) were collected. Amelogenin and 19 STR loci (D19S433, D5S818, D21S11, D18S51, D6S1043, D3S1358, D13S317, D7S820, D16S539, CSF1PO, PentaD, vWA, D8S1179, TPOX, PentaE, TH01, D12S391, D2S1338 and FGA) were tested for genotypes and examined for method identity, sensitivity, And the applicability of samples. Results The five-color fluorescent labeling multiplex amplification assay could classify the selected 20 loci and the results were stable and accurate with good balance and no heterosis. The population survey showed that the cumulative personal recognition rate and cumulative non-parent exclusion rate were 0.999 999 999 999 999 999 999 and 0.999 999 99; sensitivity of 125 pg, species-specific high, the actual case of high rate of successful biopsy. Conclusion The five-color fluorescent labeling multiplex amplification detection system indicators can reach the level of the current commercial kit, which has important value of forensic applications.