背景:大规模平行测序(MPS)允许从短串联重复(STR)分析中检索更多的信息,不仅将它们扩展到大小,正如已经在毛细管电泳(CE)中执行的那样,还有序列。MPS需要不断开发和验证分析参数,以确保STR的基因分型结果与CE获得的结果一致。鉴于Y-STRs作为常染色体STRs分析的补充标记的使用频率增加,迫切需要验证CE和MPS分析之间分型结果的一致性。
结果:使用Yfiler™PlusPCR扩增试剂盒和ForenSeq™DNASignaturePrep试剂盒对从125个无关男性唾液样本中提取的DNA进行基因分型。由SeqStudio™遗传分析仪HID和MiSeq™FGx法医基因组学系统分析,分别。对于每个共享的Y-STR,等位基因名称,每个基因座的基于长度和序列的等位基因数量,口吃百分比,筛选了多拷贝Y-STR的基因座内平衡。
结论:尽管在常规分析中应用MPS技术的法医遗传学实验室数量很少,并且不允许对MPS局限性进行全面评估,这项比较研究强调了尽管产生了大量原始数据,但MPS仍能够产生可靠的配置文件.
BACKGROUND: Massively Parallel Sequencing (MPS) allowed an increased number of information to be retrieved from short tandem repeat (STR) analysis, expanding them not only to the size, as already performed in Capillary Electrophoresis (CE), but also to the sequence. MPS requires constant development and validation of the analytical parameters to ensure that the genotyping results of STRs correspond to those obtained by CE. Given the increased frequency of usage of Y-STRs as supplementary markers to the autosomal STRs analysis, it is urgent to validate the concordance of the typing results between CE and MPS analyses.
RESULTS: DNA extracted from 125 saliva samples of unrelated males was genotyped using Yfiler™ Plus PCR Amplification Kit and ForenSeq™ DNA Signature Prep Kit, which were analyzed by SeqStudio™ Genetic Analyzer for HID and MiSeq™ FGx Forensic Genomics System, respectively. For each shared Y-STR, allele designation, number of length- and sequence-based alleles per locus, stutter percentage, and the intra-locus balance of multicopy Y-STRs were screened.
CONCLUSIONS: Although the number of forensic genetics laboratories that are applying the MPS technique in routine analysis is small and does not allow a global assessment of MPS limitations, this comparative study highlights the ability of MPS to produce reliable profiles despite the generation of large amounts of raw data.