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Isolating DNA from sexual assault cases: a comparison of standard methods with a nuclease-based approach

机译：从性侵犯案件中分离DNA：标准方法与基于核酸酶的方法的比较

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Background Profiling sperm DNA present on vaginal swabs taken from rape victims often contributes to identifying and incarcerating rapists. Large amounts of the victim’s epithelial cells contaminate the sperm present on swabs, however, and complicate this process. The standard method for obtaining relatively pure sperm DNA from a vaginal swab is to digest the epithelial cells with Proteinase K in order to solubilize the victim’s DNA, and to then physically separate the soluble DNA from the intact sperm by pelleting the sperm, removing the victim’s fraction, and repeatedly washing the sperm pellet. An alternative approach that does not require washing steps is to digest with Proteinase K, pellet the sperm, remove the victim’s fraction, and then digest the residual victim’s DNA with a nuclease. Methods The nuclease approach has been commercialized in a product, the Erase Sperm Isolation Kit (PTC Labs, Columbia, MO, USA), and five crime laboratories have tested it on semen-spiked female buccal swabs in a direct comparison with their standard methods. Comparisons have also been performed on timed post-coital vaginal swabs and evidence collected from sexual assault cases. Results For the semen-spiked buccal swabs, Erase outperformed the standard methods in all five laboratories and in most cases was able to provide a clean male profile from buccal swabs spiked with only 1,500 sperm. The vaginal swabs taken after consensual sex and the evidence collected from rape victims showed a similar pattern of Erase providing superior profiles. Conclusions In all samples tested, STR profiles of the male DNA fractions obtained with Erase were as good as or better than those obtained using the standard methods.
机译：背景技术从强奸受害者身上获取的阴道拭子上存在的精子DNA剖析通常有助于识别和监禁强奸犯。但是，受害者的大量上皮细胞会污染拭子上的精子，并使这一过程复杂化。从阴道拭子中获得相对纯净的精子DNA的标准方法是用蛋白酶K消化上皮细胞，以溶解受害人的DNA，然后通过沉淀精子，去除受害人的精子，从完整的精子中物理分离出可溶性DNA。分离，并反复洗涤精子沉淀。另一种不需要洗涤步骤的方法是用蛋白酶K消化，沉淀精子，去除受害者的部分，然后用核酸酶消化残留的受害人的DNA。方法核酸酶方法已在一种产品“擦除精子分离试剂盒”（PTC Labs，哥伦比亚，密苏里州，美国）中商业化，并且五个犯罪实验室已在精液加标的女性口腔拭子上对其进行了测试，并与标准方法进行了直接比较。还对定时性交后阴道拭子进行了比较，并从性侵犯案件中收集了证据。结果对于精液加标的口腔拭子，Erase在所有五个实验室中均优于标准方法，并且在大多数情况下，仅加标有1,500精子的颊拭子能够提供干净的男性特征。自愿性交后采集的阴道拭子和从强奸案受害者那里收集的证据显示出类似的擦除方式，具有出色的特征。结论在所有测试样品中，用Erase获得的雄性DNA片段的STR图谱与使用标准方法获得的STR图谱一样好或更好。

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