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Differential DNA extraction of challenging simulated sexual-assault samples: a Swiss collaborative study

机译：一项具有挑战性的模拟性侵犯样品的差异DNA提取：一项瑞士合作研究

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In sexual-assault cases, autosomal DNA analysis of gynecological swabs is a challenge, as the presence of a large quantity of female material may prevent detection of the male DNA. A solution to this problem is differential DNA extraction, but there is no established best practice for this. We decided to test the efficacy of a number of different protocols on simulated casework samples. Four difficult samples were sent to the nine Swiss laboratories active in forensic genetics. In each laboratory, staff used their routine protocols to separate the epithelial-cell fraction, enriched with the non-sperm DNA, from the sperm fraction. DNA extracts were then sent to the organizing laboratory for analysis. Estimates of male:female DNA ratio without differential DNA extraction ranged from 1:38 to 1:339, depending on the semen used to prepare the samples. After differential DNA extraction, most of the ratios ranged from 1:12 to 9:1, allowing detection of the male DNA. Compared with direct DNA extraction, cell separation resulted in losses of 94-98% of the male DNA. As expected, more male DNA was generally present in the sperm than in the epithelial-cell fraction. However, for about 30% of the samples, the reverse trend was seen. The recovery of male and female DNA was highly variable, depending on the laboratory involved. An experimental design similar to the one used in this study may be of assistance for local protocol testing and improvement.
机译：在性侵犯案件中，妇科拭子的常染色体DNA分析是一个挑战，因为大量雌性物质的存在可能会阻止雄性DNA的检测。解决该问题的方法是差异DNA提取，但是目前尚无最佳实践。我们决定在模拟案例工作样本上测试多种协议的功效。将四个困难样品发送到活跃在法医遗传学领域的九个瑞士实验室。在每个实验室中，工作人员使用常规方案从精子级分中分离出富含非精子DNA的上皮细胞级分。然后将DNA提取物送到组织实验室进行分析。不使用差异DNA提取方法估算的雄性：雌性DNA比例范围为1:38至1：339，具体取决于用于制备样品的精液。差异DNA提取后，大多数比率范围为1:12至9：1，从而可以检测雄性DNA。与直接DNA提取相比，细胞分离导致男性DNA损失94-98％。如预期的那样，精子中普遍存在的雄性DNA数量要多于上皮细胞部分。但是，对于大约30％的样本，可以看到相反的趋势。男性和女性DNA的回收率差异很大，这取决于所涉及的实验室。与本研究中使用的实验设计相似的实验设计可能有助于本地协议的测试和改进。

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