Objectives: To evaluate the ability of the ForenSeqTM DNA Signature Prep kit (ForenSeq kit) in analyzing the sequence information of STRs in Zhejiang She ethnic group and its forensic application efficacy.
Methods: A total of 50 Zhejiang She ethnic group samples were sequenced with the ForenSeq kit on the MiSeq FGx platform. The data was analyzed using ForenSeqTM universal analysis software to obtain the motif structure and flank regions of the 58 STRs, then compared with PCR-CE typing results to test the consistency. At last, the allele frequency and population genetic parameters were calculated.
Results: A total of 448 sequence polymorphic alleles were detected in 50 samples of Zhejiang She ethnic group. Compared with fragment length polymorphism detected by PCR-CE, 82 alleles were increased by MPS detection based on ForenSeq kit, and 7 SNPs variation were detected in the flanking regions of 6 loci. The 22 male individuals were genotyped, and total 19 haplotypes were detected in 24 Y chromosome STRs of these 22 males. The cumulative discrimination power of the 27 autosomal STRs was 1-8.87×10-30, the cumulative probability of exclusion of duo-testing was 0.999 999 962 640 657, the cumulative probability of exclusion of trios-testing was 0.999 999 999 999 633.
Conclusions: Based on MPS typing technology, using the ForenSeq kit greatly improves the detection efficiency. In addition, the 58 STRs have good genetic polymorphisms in Zhejiang She ethnic group, which are suitable for individual identification and paternity identification in forensic application.
目的: 评估ForenSeqTM DNA Signature Prep试剂盒（以下简称ForenSeq试剂盒）对浙江畲族人群STR序列的解读能力及法医学应用效能。方法: 应用MiSeq FGx法医基因组学系统和ForenSeq试剂盒对50例浙江畲族人群样本进行大规模平行测序（massively parallel sequencing，MPS）检测，采用ForenSeqTM通用分析软件分析测序数据，获取58个STR基因座的分型结果及序列结构信息。将MPS与PCR-CE技术的分型结果进行一致性比对，并计算等位基因频率和群体遗传学参数。结果: 在50例浙江畲族人群样本中，共观察到448种序列多态性等位基因，与PCR-CE技术检测的片段长度多态性结果相比，基于ForenSeq试剂盒的MPS检测增加了82个等位基因，在6个基因座的侧翼区共发现7个SNP变异。22例男性的24个Y染色体STR基因座中共检测到19种单倍型。27个常染色体STR基因座的累积个体识别率为1-8.87×10-30，累积二联体非父排除率为0.999 999 962 640 657，累积三联体非父排除率为0.999 999 999 999 633。结论: 基于MiSeq FGx法医基因组学系统使用ForenSeq试剂盒进行MPS检测，极大提高了检测效能。58个STR基因座在所调查的浙江畲族人群中具有良好的遗传多态性，可应用于法医学个体识别和亲权鉴定。.
Keywords: ForenSeq TM DNA Signature Prep kit; She ethnic group; Zhejiang; forensic genetics; massively parallel sequencing; polymorphism, genetic; short tandem repeat (STR).