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Abstract：
With the continuous development of massively parallel sequencing (MPS), increasing numbers of laboratories have utilized this method for forensic genomic analyses. When sequencing common short tandem repeats (STRs), MPS does have many advantages over the length-based genotyping method that uses traditional capillary electrophoresis (CE) technology. The Precision ID GlobalFiler™ NGS STR Panel v2 was recently released to simultaneously target 31 autosomal STRs (20 expanded Combined DNA Index System (CODIS) core loci and 11 non-CODIS loci) and 4 gender determination loci (Amelogenin, DYS391, SRY and Y-indel (rs2032678)) with the Ion S5™ System. In the current study, we performed a preliminary validation for this novel MPS-STR panel that included the following analyses: repeatability, concordance, stutter and balance, sensitivity, case-type sample testing, stability, mixture and a population investigation. Complete and reliable profiles were obtained using 125 pg of positive control DNA. The commonly encountered types of case samples and artificial mixtures with ratios of 1:1, 1:3 and 3:1 were also fully genotyped. Additional allele sequence variations were detected in samples from 50 unrelated individuals, and subsequently, an increased power of discrimination and power of exclusion were achieved. However, the average depth of coverage (DoC) of the Penta D locus was detected to be dramatically lower than those of other loci, which caused an interlocus imbalance; this could be one of the reasons for the intralocus imbalance of this locus and the 0.18% inconsistent results in the concordance study. Although certain flaws were observed, the informative metrics, including the DoC, sequence coverage ratios (SCRs) and heterozygote balance (Hb), of the novel MPS multiplex in our study were sufficient for reliable sequencing results that were 99.61% in concordance with the capillary electrophoresis (CE) results. In general, the Precision ID GlobalFiler™ NGS STR Panel v2 was demonstrated to be sensitive, reliable and robust and could be a powerful tool for human identification and kinship analyses. Additionally, we look forward to its updated version.
摘要：
随着大规模并行测序(MPS)技术的不断发展,越来越多的实验室将这种方法用于法医基因组分析。对常见的短串联重复序列(STR)进行测序时，与使用传统毛细管电泳（CE）技术的基于长度的基因分型方法相比，MPS确实具有许多优势。PrecisionIDGlobalFiler™NGSSTR面板v2最近发布，同时针对31个常染色体STR（20个扩展的联合DNA索引系统（CODIS）核心基因座和11个非CODIS基因座）和4个性别确定基因座（Amelogenin，DYS391、SRY和Y-indel(rs2032678)与IonS5™系统配合使用。在目前的研究中,我们对这个新的MPS-STR小组进行了初步验证，包括以下分析：可重复性，和谐，口吃和平衡，灵敏度,案例类型样本测试，稳定性,混合和人口调查。使用125μg阳性对照DNA获得完整和可靠的图谱。还对比例为1：1、1：3和3：1的病例样本和人工混合物的常见类型进行了完全基因分型。在50个无关个体的样本中检测到其他等位基因序列变异，随后,增加了歧视和排斥的力量。然而,检测到PentaD基因座的平均覆盖深度（DoC）显着低于其他基因座，导致基因座间失衡；这可能是该基因座内部不平衡以及一致性研究中0.18％不一致结果的原因之一。尽管观察到某些缺陷，信息度量，包括DoC,序列覆盖率(SCR)和杂合子平衡(Hb)，在我们的研究中,新的MPS多重序列足以获得与毛细管电泳(CE)结果一致的99.61%的可靠测序结果.总的来说,PrecisionIDGlobalFiler™NGSSTR面板v2被证明是敏感的，可靠和健壮，可能是人类识别和亲属关系分析的强大工具。此外,我们期待它的更新版本。
