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第二次世界大战尸骸的岩骨和掌骨的核 DNA 产量和 STR 分型成功率比较 III.

Comparison of nuclear DNA yield and STR typing success in Second World War petrous bones and metacarpals III.

机构信息

Institute of Forensic Medicine, Faculty of Medicine, University of Ljubljana, Korytkova 2, 1000 Ljubljana, Slovenia.

Faculty of Tourism Studies - Turistica, University of Primorska, Portorož, Slovenia.

出版信息

Forensic Sci Int Genet. 2021 Nov;55:102578. doi: 10.1016/j.fsigen.2021.102578. Epub 2021 Aug 18.

Abstract

DNA yield and STR typing success differ among skeletal element types and within individual bones. Consequently, it is necessary to identify the skeletal elements and their intra-skeletal parts that will most likely yield utilizable and informative endogenous DNA for human identification of skeletal remains. The petrous portion of the temporal bone has been shown to be the most suitable skeletal part for sampling archaeological skeletons, and it has also been used successfully in some forensic cases. When all representative bone types were analyzed for three complete Second World War skeletons, metatarsals and metacarpals yielded more DNA than petrous bones (which generated full profiles even if the DNA yield was lower) and, among almost 200 Second World War metatarsals and metacarpals analyzed, metacarpals III were found to be the highest-yielding bones. To further improve the sampling strategy in DNA analysis of aged skeletal remains, a comparison between 26 petrous bones and 30 metacarpals III from Second World War skeletons was made considering intra-bone DNA yield variability. In metacarpals III only epiphyses were sampled, and in petrous bones only the dense part within the otic capsule was used. To exclude the influence of taphonomic issues as much as possible, petrous bones and metacarpals III from a single Second World War mass grave were examined. The difference between petrous bones and metacarpals III was explored by measuring nuclear DNA yield and success of STR typing. After cleaning the samples, full demineralization extraction was used to decalcify 0.5 g of powdered bone. PowerQuant (Promega) was used to determine DNA content and DNA degradation rates, and STR typing was performed using the PowerPlex ESI 17 Fast System (Promega). Metacarpals III produced the same DNA yields and STR typing success as petrous bones with no intra-individual difference observed in concentration of DNA, degradation rate, percentage of successfully amplified alleles, and intensity of electrophoretic signals. Sampling and processing of metacarpal III epiphyses is consequently recommended for genetic identification of highly degraded skeletal remains in routine forensic casework and in buried non-commingled aged skeletal remains whenever metacarpals III are preserved. Metacarpals III are easy to sample and less prone to contamination with modern DNA because no saw is needed for sampling in comparison to the petrous portion of the temporal bone. The data obtained in this study further improve the sampling strategy for genetic identification of Second World War skeletal remains in Slovenia.

摘要

DNA 产量和 STR 分型成功率因骨骼类型而异,甚至在单个骨骼内也存在差异。因此,有必要确定最有可能为骨骼遗骸的人类身份识别提供可用和有信息价值的内源性 DNA 的骨骼成分及其内部骨骼部分。颞骨的岩部已被证明是用于采样考古骨骼的最合适的骨骼部分,并且在一些法医案例中也已成功使用。当对三具完整的二战骨骼进行所有代表性骨骼类型分析时,跖骨和掌骨的 DNA 产量高于岩骨(即使 DNA 产量较低,也能生成完整的图谱),在分析的近 200 块二战跖骨和掌骨中,发现掌骨 III 的产量最高。为了进一步改进老化骨骼遗骸 DNA 分析中的采样策略,对来自二战骨骼的 26 块岩骨和 30 块掌骨 III 进行了比较,考虑了骨骼内 DNA 产量的可变性。在掌骨 III 中仅采集了骺,而在岩骨中仅使用了耳囊内的致密部分。为了尽可能排除埋藏学问题的影响,对来自二战乱葬岗的单个岩骨和掌骨 III 进行了检查。通过测量核 DNA 产量和 STR 分型成功率来探索岩骨和掌骨 III 之间的差异。在对样本进行清洁后,使用全脱矿化提取法对 0.5 克粉末状骨骼进行脱钙。使用 PowerQuant(Promega)来确定 DNA 含量和 DNA 降解率,并使用 PowerPlex ESI 17 Fast System(Promega)进行 STR 分型。掌骨 III 的 DNA 产量和 STR 分型成功率与岩骨相同,没有观察到个体内 DNA 浓度、降解率、成功扩增等位基因的百分比和电泳信号强度的差异。因此,建议在常规法医工作中以及在保存掌骨 III 的情况下,对高度降解的骨骼遗骸进行遗传鉴定时,采样和处理掌骨 III 骺。与颞骨岩部相比,采样时不需要锯子,因此掌骨 III 易于采样且不易受到现代 DNA 的污染。本研究获得的数据进一步改进了在斯洛文尼亚对二战骨骼遗骸进行遗传鉴定的采样策略。

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