Institute of Legal Medicine, Medical University of Innsbruck, Innsbruck, Austria.
Center for Human Identification, University of North Texas Health Science Center, TX, USA.
Forensic Sci Int Genet. 2019 Sep;42:244-251. doi: 10.1016/j.fsigen.2019.07.013. Epub 2019 Jul 23.
The emergence of Massively Parallel Sequencing technologies enabled the analysis of full mitochondrial (mt)DNA sequences from forensically relevant samples that have, so far, only been typed in the control region or its hypervariable segments. In this study, we evaluated the performance of a commercially available multiplex-PCR-based assay, the Precision ID mtDNA Whole Genome Panel (Thermo Fisher Scientific), for the amplification and sequencing of the entire mitochondrial genome (mitogenome) from even degraded forensic specimens. For this purpose, more than 500 samples from 24 different populations were selected to cover the vast majority of established superhaplogroups. These are known to harbor different signature sequence motifs corresponding to their phylogenetic background that could have an effect on primer binding and, thus, could limit a broad application of this molecular genetic tool. The selected samples derived from various forensically relevant tissue sources and were DNA extracted using different methods. We evaluated sequence concordance and heteroplasmy detection and compared the findings to conventional Sanger sequencing as well as an orthogonal MPS platform. We discuss advantages and limitations of this approach with respect to forensic genetic workflow and analytical requirements.
高通量测序技术的出现使得能够从法医相关样本中分析完整的线粒体（mt）DNA 序列，而这些样本迄今为止仅在控制区域或其高变区进行了分型。在这项研究中，我们评估了一种商业上可获得的基于多重 PCR 的检测方法，即 Precision ID mtDNA 全基因组面板（Thermo Fisher Scientific），用于从即使降解的法医样本中扩增和测序整个线粒体基因组（mitogenome）。为此，从 24 个不同群体中选择了 500 多个样本，以涵盖绝大多数已建立的超级单倍群。这些单倍群已知含有与其进化背景相对应的不同特征序列基序，这可能会影响引物结合，从而限制这种分子遗传学工具的广泛应用。所选样本来自各种法医相关组织来源，并使用不同的方法提取 DNA。我们评估了序列一致性和异质性检测，并将结果与传统的 Sanger 测序以及正交 MPS 平台进行了比较。我们讨论了这种方法在法医遗传学工作流程和分析要求方面的优缺点。
