Institute of Legal Medicine, Medical University of Innsbruck, 6020 Innsbruck, Tirol, Austria.
BIOMICs Research Group, Lascaray Research Center, University of the Basque Country, 01006 Vitoria-Gasteiz, Basque Country, Spain.
Genes (Basel). 2020 Jun 26;11(6):708. doi: 10.3390/genes11060708.
The study of DNA to predict externally visible characteristics (EVCs) and the biogeographical ancestry (BGA) from unknown samples is gaining relevance in forensic genetics. Technical developments in Massively Parallel Sequencing (MPS) enable the simultaneous analysis of hundreds of DNA markers, which improves successful Forensic DNA Phenotyping (FDP). The EU-funded VISAGE (VISible Attributes through GEnomics) Consortium has developed various targeted MPS-based lab tools to apply FDP in routine forensic analyses. Here, we present an evaluation of the VISAGE Basic tool for appearance and ancestry prediction based on PowerSeq chemistry (Promega) on a MiSeq FGx System (Illumina). The panel consists of 153 single nucleotide polymorphisms (SNPs) that provide information about EVCs (41 SNPs for eye, hair and skin color from HIrisPlex-S) and continental BGA (115 SNPs; three overlap with the EVCs SNP set). The assay was evaluated for sensitivity, repeatability and genotyping concordance, as well as its performance with casework-type samples. This targeted MPS assay provided complete genotypes at all 153 SNPs down to 125 pg of input DNA and 99.67% correct genotypes at 50 pg. It was robust in terms of repeatability and concordance and provided useful results with casework-type samples. The results suggest that this MPS assay is a useful tool for basic appearance and ancestry prediction in forensic genetics for users interested in applying PowerSeq chemistry and MiSeq for this purpose.
研究 DNA 以从未知样本中预测外部可见特征 (EVC) 和生物地理祖先 (BGA) 在法医学中越来越受到关注。大规模平行测序 (MPS) 的技术发展使同时分析数百个 DNA 标记成为可能，从而提高了法医 DNA 表型预测 (FDP) 的成功率。由欧盟资助的 VISAGE (通过基因组学实现可见属性) 联盟开发了各种基于靶向 MPS 的实验室工具，以将 FDP 应用于常规法医分析。在这里，我们根据 PowerSeq 化学 (Promega) 在 MiSeq FGx 系统 (Illumina) 上评估了 VISAGE Basic 工具在外观和祖先预测方面的性能。该面板由 153 个单核苷酸多态性 (SNP) 组成，可提供有关 EVC（HIrisPlex-S 中的 41 个 SNP 用于眼睛、头发和肤色）和大陆 BGA（115 个 SNP；3 个与 EVC 重叠 SNP 集）的信息。该测定法针对灵敏度、重复性和基因分型一致性以及与案例样本的性能进行了评估。这种靶向 MPS 测定法在 125 pg 输入 DNA 时可提供所有 153 个 SNP 的完整基因型，在 50 pg 时可提供 99.67%的正确基因型。它在重复性和一致性方面具有稳健性，并为案例样本提供了有用的结果。结果表明，对于有兴趣在法医遗传学中应用 PowerSeq 化学和 MiSeq 的用户来说，这种 MPS 测定法是一种用于基本外观和祖先预测的有用工具。
