High-throughput DNA extraction of forensic adhesive tapes
High-throughput DNA extraction of forensic adhesive tapes
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DOI：
10.1016/j.fsigen.2016.06.004
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发表时间：
2016-09-01
影响因子：
3.1
通讯作者：
Hedman, Johannes
中科院分区：
文献类型：
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作者：
Forsberg, Christina;Jansson, Linda;Hedman, Johannes
自21世纪初引入以来，胶带粘取法已成为法医DNA分析中一种成熟的取样方法。取样快速且简单，但由于胶带的“粘性”、刚性和尺寸，后续的DNA提取更具挑战性。我们已经开发、验证并实施了一种用于胶带的简单高效的直接裂解DNA提取方案，该方案所需的人工操作较少。该方法使用Chelex beads，并应用于SceneSafe FAST胶带。这种直接裂解方案在Automate Express上提供的平均DNA产量高于PrepFiler Express BTA，尽管在使用以受控方式穿着的衣物作为参考材料时差异不显著（对于T恤和有领衬衫，p值分别为0.13和0.34）。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0.01 ng/mL的提取物比例从71%提高到了76%。除了与之前的方法相比提供更高的DNA产量外，所开发的直接裂解方案的引入还将人工工作量减少了一半，并使实验室中胶带的潜在通量翻倍。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具有成本效益的替代方案。（C）2016作者。由爱思唯尔爱尔兰有限公司出版。
Tape-lifting has since its introduction in the early 20000s 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 0.01 ng/mL 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. (C) 2016 The Authors. Published by Elsevier Ireland Ltd.