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爆炸后简易爆炸装置样本经荧光可视化和直接 PCR 成功进行 STR 扩增。

Successful STR amplification of post-blast IED samples by fluorescent visualisation and direct PCR.

机构信息

College of Science & Engineering, Flinders University, Adelaide, Australia.

College of Science & Engineering, Flinders University, Adelaide, Australia.

出版信息

Forensic Sci Int Genet. 2020 May;46:102256. doi: 10.1016/j.fsigen.2020.102256. Epub 2020 Jan 29.

Abstract

Improvised explosive devices (IEDs) present a number of challenges in terms of the generation of forensically relevant information. Inhibition to PCR from sub-optimal sample types as well as from specific substrates has historically meant that extraction prior to PCR has been required. Improvements to STR kit buffers lead to the successful introduction of direct PCR to the analysis of IED-relevant samples, however none of these samples have been exposed to detonations. This study presents data to support the use of direct PCR in the analysis of IED components post-detonation. VeriFiler™ Plus generated informative profiles, containing ≥ 12 autosomal alleles, from samples touched for a maximum of 15 s that were then exposed to a detonation from plastic explosive placed as close as 100 mm. Of the 37 recovered touched items or fragments, 28 contained autosomal alleles from the donor with 18 (49 %) presenting informative profiles that matched the DNA donor. This compared with results following STR PCR post-extraction with one of 11 amplified post-detonation touch DNA samples being informative. The use of Diamond™ Nucleic Acid Dye (DD) staining and visualisation before and after detonation allowed for analysis as to cell loss or damage as a result of the detonation itself and aided in the triaging of samples to be selected for DNA profiling. This is the first record of cellular visualisation and comparison before and after detonation with accompanying STR results on a range of sample types typical of IED constituents. Following comparison of DD visualised cells and STR amplification success, chemical analysis of plastic and electrical tape samples supported substrate-specific inhibition. These data represent the first instance of informative DNA profiles being produced from post-detonation samples using direct PCR, as close as 100 mm from the charge.

摘要

简易爆炸装置 (IED) 在生成法证相关信息方面存在诸多挑战。由于次优样本类型以及特定基质对 PCR 的抑制作用,历史上一直需要在 PCR 之前进行提取。STR 试剂盒缓冲液的改进使得直接 PCR 能够成功应用于 IED 相关样本的分析,但这些样本都没有暴露于爆炸中。本研究提供的数据支持在爆炸后对 IED 成分进行直接 PCR 分析。VeriFiler Plus 试剂盒从接触时间最长为 15 秒的样本中生成了包含 12 个以上常染色体等位基因的信息丰富的图谱,这些样本随后暴露于距离最近 100 毫米的塑料炸药爆炸中。在所回收的 37 个接触物品或碎片中,有 28 个包含供体的常染色体等位基因,其中 18 个(49%)呈现出与 DNA 供体匹配的信息丰富图谱。相比之下,对提取后进行 STR PCR 的 11 个爆炸后接触 DNA 样本中的一个进行分析的结果,只有一个是信息丰富的。在爆炸前后使用 Diamond 核酸染料 (DD) 染色和可视化,可以分析爆炸本身导致的细胞丢失或损伤,并有助于对要选择进行 DNA 分析的样本进行分类。这是首次记录在案的对爆炸前后的细胞进行可视化比较,并附有一系列 IED 成分典型样本的 STR 结果。在比较 DD 可视化细胞和 STR 扩增成功率之后,对塑料和电工胶带样本的化学分析支持基质特异性抑制作用。这些数据代表了首次使用直接 PCR 从爆炸后样本中产生信息丰富的 DNA 图谱,距离爆炸源最近可达 100 毫米。

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