College of Science and Engineering, Flinders University, SA 5042, Australia.
College of Science and Engineering, Flinders University, SA 5042, Australia; Forensic Science SA, 21 Divett Place, Adelaide, SA 5000, Australia.
Forensic Sci Int Genet. 2018 Sep;36:13-19. doi: 10.1016/j.fsigen.2018.06.002. Epub 2018 Jun 2.
Direct PCR from touch DNA has a range of potential applications in the field of forensic investigation for exhibit examination that, under standard extraction methods, rarely produce informative DNA profiles. Previous studies from 'touch DNA' have focussed on fingermarks created under laboratory conditions. Here we report on successful STR DNA profiling from a range of touched items. Direct PCR, with no increase in cycle number, was performed after eight different sample types, typical of those submitted for forensic investigation, were handled by volunteers for a maximum of 15 s to deposit trace amounts of their DNA. Amplifications were performed using either GlobalFiler or Identifiler Plus following manufacturer's instructions. These two kits were chosen deliberately as many laboratories worldwide have adopted and validated them in their workflow, thus allowing for direct PCR to be incorporated within their practises easily. It was found that informative STR profiles were obtained from all eight substrates using both STR kits. Identifiler Plus out-performed GlobalFiler in terms of the percentage of alleles amplified using the direct PCR approach. Both generated informative profiles from all items and all individuals, at different rates, with Identifiler Plus being informative in a larger percentage of samples. GlobalFiler produced profiles with an average of 60% ± 24% (36 ± 15 alleles) alleles present while Identifiler Plus produced profiles with an average of 96% ± 4% (31 ± 1 alleles) alleles present. A comparison was made between the direct PCR approach and subjecting touched samples to a standard DNA extraction process, both using Identifiler. An average of 4% of profiles were informative for samples that underwent extraction with 100% being informative from the same subset of samples amplified by direct PCR. Our findings further demonstrate the success of direct PCR to enhance the STR DNA profiles from touch DNA. Further, Identifiler Plus was found to generate informative profiles more often than GlobalFiler. Direct PCR is fast, simple, and non-destructive of evidence with the ability to generate informative genetic data where standard methods are likely to fail.
直接从接触性 DNA 进行 PCR 分析在法庭科学领域具有广泛的应用潜力,因为在标准提取方法下,很少能从证物中提取到有意义的 DNA 谱。之前关于“接触性 DNA”的研究主要集中在实验室条件下产生的指纹上。在这里,我们报告了从一系列接触性物品中成功进行 STR 基因分型的结果。在志愿者处理了八种不同类型的样本(这些样本通常提交给法庭科学调查),最多接触 15 秒以留下微量 DNA 后,无需增加循环次数,直接进行 PCR 分析。扩增是按照制造商的说明使用 GlobalFiler 或 Identifiler Plus 试剂盒进行的。这两种试剂盒是特意选择的,因为全世界许多实验室已经采用并在其工作流程中验证了它们,因此可以很容易地将直接 PCR 纳入其实践。结果发现,使用这两种试剂盒都可以从所有八种基质中获得有意义的 STR 谱。Identifiler Plus 在直接 PCR 方法中扩增的等位基因百分比方面优于 GlobalFiler。两种试剂盒都以不同的速度从所有物品和所有个体中生成有意义的谱,Identifiler Plus 在更大比例的样本中具有信息性。GlobalFiler 产生的谱平均有 60%±24%(36±15 个等位基因)存在的等位基因,而 Identifiler Plus 产生的谱平均有 96%±4%(31±1 个等位基因)存在的等位基因。比较了直接 PCR 方法和对接触性样本进行标准 DNA 提取处理的方法,两者都使用 Identifiler。经过提取处理的样本中,有 4%的谱是有意义的,而直接 PCR 方法扩增的相同样本子集的 100%谱是有意义的。我们的研究结果进一步证明了直接 PCR 能够增强接触性 DNA 的 STR DNA 谱。此外,Identifiler Plus 比 GlobalFiler 更常产生有意义的谱。直接 PCR 快速、简单、对证据无损害,并且能够在标准方法可能失败的情况下生成有意义的遗传数据。