Institute of Forensic Medicine, Faculty of Medicine, University of Ljubljana, Korytkova 2, 1000, Ljubljana, Slovenia.
Faculty of Tourism Studies - Turistica, University of Primorska, Portorož, Slovenia.
Int J Legal Med. 2020 Sep;134(5):1629-1638. doi: 10.1007/s00414-020-02289-x. Epub 2020 May 6.
For identification of badly preserved cadavers, only a few tissues can be used as a source of DNA, mostly bones and teeth, from which sampling and DNA extraction are difficult and time-consuming. In most highly decomposed remains, the nails are preserved. The aim of this study was to evaluate nails as an alternative source of DNA instead of bones and teeth in demanding routine identification cases. An automated extraction method was optimized on nails obtained from 33 cadavers with a post-mortem interval (PMI) up to 5 years. The commercially available EZ1 Investigator Kit (Qiagen) was used for extraction, and the G2 buffer included in the kit was replaced with TN buffer, and DTT was added for digestion of 5 mg of nail. The DNA was purified in a Biorobot EZ1 device (Qiagen), quantified using the PowerQuant System (Promega), and STR typing was performed with the NGM kit (TFS). From 0.3 to 270 μg DNA/g of nail was obtained from the samples analyzed, with an average yield of 36 μg DNA/g of nail. Full STR profiles were obtained from all nails except one. The optimized extraction method proved to be fast and highly efficient in the removal of PCR inhibitors, and it yields high amounts of DNA for successful STR typing. Nails were implemented as the primary sample type for obtaining DNA from highly decomposed and partially skeletonized cadavers in routine forensic identification cases in our laboratory.
为了识别保存状况不佳的尸体,只能使用少数组织作为 DNA 的来源,主要是骨头和牙齿,从这些组织中采样和提取 DNA 既困难又耗时。在大多数高度腐烂的遗骸中,指甲得以保存。本研究旨在评估指甲作为一种替代骨头和牙齿的 DNA 来源,用于在要求苛刻的常规鉴定案例中。我们优化了一种自动化提取方法,用于从 33 具死后间隔时间(PMI)长达 5 年的尸体上获得的指甲。使用市售的 EZ1 Investigator 试剂盒(Qiagen)进行提取,并将试剂盒中的 G2 缓冲液替换为 TN 缓冲液,添加 DTT 以消化 5mg 指甲。在 Biorobot EZ1 设备(Qiagen)中纯化 DNA,使用 PowerQuant 系统(Promega)定量,并使用 NGM 试剂盒(TFS)进行 STR 分型。从分析的样本中获得了 0.3 至 270μg/g 指甲的 DNA,平均每克指甲获得 36μg DNA。除了一个样本之外,所有指甲都获得了完整的 STR 图谱。优化的提取方法被证明在去除 PCR 抑制剂方面快速且高效,并且能够产生大量的 DNA,从而成功进行 STR 分型。在我们实验室的常规法医鉴定案例中,指甲被用作从高度腐烂和部分骨骼化的尸体中获取 DNA 的主要样本类型。