Forsberg Christina, Jansson Linda, Ansell Ricky, Hedman Johannes
Swedish National Forensic Centre, Linköping, Sweden.
Applied Microbiology, Department of Chemistry, Lund University, Lund, Sweden.
Forensic Sci Int Genet. 2016 Sep;24:158-163. doi: 10.1016/j.fsigen.2016.06.004. Epub 2016 Jun 7.
Tape-lifting has since its introduction in the early 2000's become a well-established sampling method in forensic DNA analysis. Sampling is quick and straightforward while the following DNA extraction is more challenging due to the "stickiness", rigidity and size of the tape. We have developed, validated and implemented a simple and efficient direct lysis DNA extraction protocol for adhesive tapes that requires limited manual labour. The method uses Chelex beads and is applied with SceneSafe FAST tape. This direct lysis protocol provided higher mean DNA yields than PrepFiler Express BTA on Automate Express, although the differences were not significant when using clothes worn in a controlled fashion as reference material (p=0.13 and p=0.34 for T-shirts and button-down shirts, respectively). Through in-house validation we show that the method is fit-for-purpose for application in casework, as it provides high DNA yields and amplifiability, as well as good reproducibility and DNA extract stability. After implementation in casework, the proportion of extracts with DNA concentrations above 0.01ng/μL increased from 71% to 76%. Apart from providing higher DNA yields compared with the previous method, the introduction of the developed direct lysis protocol also reduced the amount of manual labour by half and doubled the potential throughput for tapes at the laboratory. Generally, simplified manual protocols can serve as a cost-effective alternative to sophisticated automation solutions when the aim is to enable high-throughput DNA extraction of complex crime scene samples.
自21世纪初引入以来,胶带提取已成为法医DNA分析中一种成熟的采样方法。采样快速且直接,但由于胶带的“粘性”、硬度和尺寸,后续的DNA提取更具挑战性。我们开发、验证并实施了一种简单高效的直接裂解DNA提取方案,用于胶带,所需的人工操作有限。该方法使用Chelex磁珠,并应用于SceneSafe FAST胶带。尽管在以受控方式穿着的衣物作为参考材料时(T恤和扣领衬衫的p值分别为0.13和0.34),直接裂解方案的平均DNA产量高于Automate Express上的PrepFiler Express BTA,但差异并不显著。通过内部验证,我们表明该方法适用于实际案件工作,因为它提供了高DNA产量和可扩增性,以及良好的重现性和DNA提取物稳定性。在实际案件工作中实施后,DNA浓度高于0.01ng/μL的提取物比例从71%增加到了76%。与之前的方法相比,除了提供更高的DNA产量外,所开发的直接裂解方案的引入还将人工操作量减少了一半,并使实验室胶带的潜在通量增加了一倍。一般来说,当目标是实现复杂犯罪现场样本的高通量DNA提取时,简化的手动方案可以作为复杂自动化解决方案的一种经济高效的替代方案。