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首页> 外文期刊>Science & justice: journal of the Forensic Science Society >Recovery of human DNA profiles from poached deer remains part 2: Improved recovery protocol without the need for LCN analysis
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Recovery of human DNA profiles from poached deer remains part 2: Improved recovery protocol without the need for LCN analysis

机译：从水煮鹿中恢复人类DNA的特征仍然是第2部分：无需LCN分析就可以改进恢复方案

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Although poaching is a common wildlife crime, the high and prohibitive cost of specialised animal testing means that many cases are left un-investigated. We previously described a novel approach to wildlife crime investigation that looked at the identification of human DNA on poached animal remains (Tobe, Govan and Welch, 2011). Human DNA was successfully isolated and amplified from simulated poaching incidents, however a low template protocol was required which made this method unsuitable for use in many laboratories. We now report on an optimised recovery and amplification protocol which removes the need for low template analysis.Samples from 10 deer (40 samples total - one from each leg) analysed in the original study were re-analysed in the current study with an additional 11 deer samples. Four samples analysed using Chelex did not show any results and a new method was devised whereby the available DNA was concentrated. By combining the DNA extracts from all tapings of the same deer remains followed by concentration, the recovered quantity of human DNA was found to be 29.5pg±43.2pg, 31× greater than the previous study. The use of the Investigator Decaplex SE (QIAGEN) STR kit provided better results in the form of more complete profiles than did the AmpF?STR? SGM Plus? kit at 30cycles (Applied Biosystems). Re-analysis of the samples from the initial study using the new, optimised protocol resulted in an average increase of 18% of recovered alleles. Over 17 samples, 71% of the samples analysed using the optimised protocol showed sufficient amplification for comparison to a reference profile and gave match probabilities ranging from 7.7690×10-05 to 2.2706×10-14.The removal of low template analysis means this optimised method provides evidence of high probative value and is suitable for immediate use in forensic laboratories. All methods and techniques used are standard and are compatible with current SOPs. As no high cost non-human DNA analysis is required the overall process is no more expensive than the investigation of other volume crime samples. The technique is suitable for immediate use in poaching incidents.
机译：尽管偷猎是一种常见的野生动植物犯罪，但是进行专门的动物检测的高昂费用高昂，这意味着许多案件没有得到调查。我们之前描述了一种新颖的野生动植物犯罪调查方法，该方法着眼于水煮动物遗体上人类DNA的鉴定（Tobe，Govan和Welch，2011年）。已成功地从模拟偷猎事件中分离并扩增了人类DNA，但是需要低模板规程，这使得该方法不适用于许多实验室。现在我们报告了一种优化的回收和扩增方案，该方案无需进行低模板分析。在当前研究中，重新分析了原始研究中分析的10头鹿（共40个样品-每条腿各一个）的样品。鹿样品。使用Chelex分析的四个样品均未显示任何结果，因此设计了一种新方法来浓缩可用的DNA。通过合并来自相同鹿残骸的所有录音带的DNA提取物，然后进行浓缩，发现人类DNA的回收量为29.5pg±43.2pg，比以前的研究大31倍。与AmpF？STR？相比，使用Investigator Decaplex SE（QIAGEN）STR试剂盒可以提供更完善的结果，从而提供更好的结果。 SGM Plus？ 30个循环的试剂盒（Applied Biosystems）。使用新的优化方案对来自初始研究的样品进行重新分析，结果平均回收的等位基因增加了18％。在超过17个样品中，使用优化方案分析的样品中有71％显示出足够的扩增以与参考谱图进行比较，并给出了7.7690×10-05至2.2706×10-14的匹配概率。该方法提供了高证明价值的证据，适合在法医实验室立即使用。所有使用的方法和技术都是标准的，并且与当前的SOP兼容。由于不需要高成本的非人类DNA分析，因此整个过程并不比调查其他大量犯罪样本更昂贵。该技术适合在偷猎事件中立即使用。

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