19个常染色体与4个Y染色体STR复合扩增体系的建立

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ABI3130XL遗传分析仪对扩增产物进行电泳，GeneMapperID3．2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0．999999999，三联体累计非父排除率达0．999999985，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。

Development of a novel multiplex PCR system with 19 autosomal STR and 4 Y-STR loci

Objective To establish a novel multiplex PCR system and assess its value in forensic application. Methods The novel system was a 5-color fluorescen dye system which contained 19 autosomal STR and 4 Y-STR loci, as well as amelogenin. The PCR products were separated on ABI 3130XL genetic analyzer and then analyzed by GeneMapper ID 3.2 software. The sensitivity, stability, reproductivity, species specificity experiments as well as contact, mixtures and degraded samples were organized, and the profiles were compared with that from the PowerPlex~ 21 System. In addition, 500 unrelated Chinese individuals from Guangdong province were tested to calculate polymorphic statistical parameters for forensic use. Results The genotyping results could be well obtained from 0.05ng to 1.0ng DNA, which were accurate and balance. The mixtures, degraded samples, contact samples were also correctly typed. The total discrimination power of the 19 autosomal STR loci was above 0. 999 999 999, and the cumulative probability of paternity exclusion for triplet cases was 0. 999 999 985. In addition, 148 different haplotypes were observed from 250 male unrelated individuals from Guangdong Han population, and the haplotype diversity was 0. 592. Conclusion The multiplex PCR system developed by us is robust and reliable, and is of high value in forensic application.