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目的建立19个常染色体STR及Amelogenin和4个Y染色体STR基因座复合扩增体系,并对其效能进行评估。方法用五色荧光标记20+4Y-STR基因座,建立同步扩增检测体系,用ABI 3130XL遗传分析仪对扩增产物进行电泳,GeneMapperID 3.2软件进行基因分型;检测体系的灵敏度、均衡性、稳定性、特异性、同一性和稳定性,并观察混合、降解及微量检材的分型情况。结果采用本文体系,DNA模板量在0.05~1.00ng时,分型准确,均衡性、特异性好;混合、降解及微量检材分型正确。该19个常染色体STR基因座的累计个人识别率大于0.999 999 999,三联体累计非父排除率达0.999 999 985,Y-STR单倍型多态性为0.592。结论本文建立的复合扩增体系分型准确,稳定,在法医学案件检验及数据库建设等方面有良好的应用前景。
Objective To establish a composite amplification system of 19 autosomal STR, Amelogenin and 4 Y chromosome STR loci and to evaluate their efficacy. Methods A multiplex PCR amplification system was established by using 20 + 4Y-STR loci. The amplification products were electrophoresed on a ABI 3130XL Genetic Analyzer. GeneMapperID 3.2 software was used for genotyping. The sensitivity, equilibrium and stability of the system were tested Sex, specificity, identity and stability, and observe the mixture, degradation and micro-sample classification. Results The system of this article was accurate, well-balanced and specific when the amount of DNA template was between 0.05 and 1.00ng. The typing, mixing, degrading and micro-typing were correct. The cumulative autosensing rate of the 19 autosomal STR loci was greater than 0.999 999 999. The triple non-parent exclusion rate was 0.999 999 985 and the Y-STR haplotype diversity was 0.592. Conclusion The composite amplification system established in this paper is accurate and stable, and it has a good application prospect in forensic case testing and database construction.