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目的建立6个Y-STR荧光复合扩增体系,评价其法医学应用价值。方法设计DYS444,GATA-A7.2,GATA-A10,DYS390,GATA-A7.1,DYS443荧光复合扩增引物,扩增总体积20μL,内含模板DNA0.5～10ng,PCR产物用3130遗传分析仪电泳,GenemapperID v 3.2分析结果,根据等位基因标准命名各等位基因,并评价该系统的特异性、准确性、均衡性、灵敏度及对混合血样的分析能力。结果当dNTP、Mg2+分别为200μmol/L、1.5mmol/L时扩增效果最佳,0.5～10ngDNA模板量均能获得较好的扩增效果,各基因座分型结果清晰,扩增均衡,特异性强,重现性好,基本满足实际应用的性能要求。对湖北汉族群体进行遗传学调查,结果GD值0.580 3～0.722 3,共检出158种单倍型,其多样性为0.995 8。结论本文6个Y-STR扩增系统分型可靠,配合常用Y-STR分型试剂盒,可进一步提高个体识别能力。
Objective To establish six Y-STR fluorescence multiplex amplification system to evaluate the forensic value. Methods The primers of DYS444, GATA-A7.2, GATA-A10, DYS390, GATA-A7.1 and DYS443 were designed and synthesized. The total volume of amplification was 20μL, containing 0.5-10ng of template DNA. The PCR products were analyzed by 3130 genetic analysis Electrophoresis, Genemapper ™ ID v 3.2 analysis of the results, named according to the allele alleles, and to evaluate the system specificity, accuracy, balance, sensitivity and analysis of mixed blood samples. Results The results showed that the best amplification effect was obtained when dNTPs and Mg2 + were 200μmol / L and 1.5mmol / L, respectively. The results showed that the amplification results were clear, Strong, reproducible, and basically meet the performance requirements of the actual application. Genetic survey of Han population in Hubei Province showed that the GD value ranged from 0.580 3 to 0.722 3. A total of 158 haplotypes were detected with a diversity of 0.995 8. Conclusion The 6 Y-STR amplification systems are reliable and can be used in combination with the commonly used Y-STR typing kit to further improve the individual identification ability.