Institute of Legal Medicine, Goethe University, University Hospital Frankfurt, Frankfurt, Germany; German Red Cross Blood Center, Institute of Transfusion Medicine and Immunohaematology, University Hospital Frankfurt, Frankfurt, Germany.
German Red Cross Blood Center, Institute of Transfusion Medicine and Immunohaematology, University Hospital Frankfurt, Frankfurt, Germany.
Forensic Sci Int. 2021 May;322:110768. doi: 10.1016/j.forsciint.2021.110768. Epub 2021 Mar 17.
Over the past years, next-generation sequencing (NGS) technologies revolutionized the possibilities in a broad range of application areas. Also in the field of forensic genetics, NGS continuously gained in importance and attentiveness. A significant number of sudden cardiac deaths (SCD) in the young is due to heritable arrhythmia syndromes emphasizing the need of examining the genetic basis in these cases also with regard to the identification of relatives and/or patients being at risk. As a result, high-throughput methods became of increasing value in molecular autopsy investigations enabling the analysis of a broad spectrum of genes. Most standard protocols are optimized for high-quality samples and frequently not directly applicable to challenging forensic sample material. In the present study, we intended to examine a comprehensive gene panel associated with SCD and inherited arrhythmogenic disorders. We compared three different hybridization-based library preparation technologies in order to implement a suitable NGS workflow for heterogeneous, forensic as well as diagnostic sample material. The results obtained indicated, that the Illumina technologies Nextera DNA Flex and TruSeq were compatible with samples exhibiting varying levels of degradation. In comparison, the TruSight method also resulted in good sequencing data, but seemed to be more dependent on DNA integrity. The preparation protocols evaluated in our study are not restricted to molecular autopsy investigations and might be helpful for and transferrable to further forensic research applications.
在过去的几年中，下一代测序（NGS）技术在广泛的应用领域中带来了革命性的变化。在法医遗传学领域，NGS 也不断受到重视。大量年轻人的突发性心脏死亡（SCD）是由于遗传性心律失常综合征引起的，这强调了在这些情况下也需要检查遗传基础，以便识别亲属和/或处于风险中的患者。因此，高通量方法在分子尸检研究中变得越来越有价值，能够分析广泛的基因。大多数标准方案都针对高质量的样本进行了优化，并且通常不能直接应用于具有挑战性的法医样本材料。在本研究中，我们旨在检查与 SCD 和遗传性心律失常障碍相关的综合基因谱。我们比较了三种不同的基于杂交的文库制备技术，以便为异质、法医和诊断样本材料实施合适的 NGS 工作流程。结果表明，Illumina 的 Nextera DNA Flex 和 TruSeq 技术与具有不同降解水平的样本兼容。相比之下，TruSight 方法也能产生良好的测序数据，但似乎更依赖于 DNA 完整性。我们研究中评估的制备方案不仅限于分子尸检研究，而且可能有助于并可转移到进一步的法医研究应用中。
