Institute of Forensic Medicine, Faculty of Medicine, University of Ljubljana, Korytkova 2, Ljubljana 1000, Slovenia.
Department of Medicine, Surgery, and Health, University of Trieste, Trieste 34137, Italy.
Forensic Sci Int. 2024 Sep;362:112184. doi: 10.1016/j.forsciint.2024.112184. Epub 2024 Aug 2.
The petrous bone contains significantly higher amounts of DNA than any other human bone. Because of highly destructive sampling and because it is not always part of the recovered remains, the need for alternative sources of DNA is important. To identify additional optimal bone types, petrous bones were compared to femurs, tali, and calcanei sampled from 66 adult skeletons from two distinct modern-era Christian cemeteries. An extraction method employing full demineralization was used to obtain DNA, real-time PCR quantification to ascertain DNA quantity and degradation, and a commercial forensic short tandem repeats (STR) PCR amplification kit to determine genetic profiles. Statistical analysis was performed to explore the differences in DNA yield, DNA degradation, and success of STR amplification. A systematic studies exploring intra-skeletal variability in DNA preservation including various excavation sites differing by time period and geographical position are rare, and the second part of the investigation was based on a comparison of both archaeological sites, which allowed us to compare the effect of different post-mortem intervals and environmental conditions on DNA preservation. The older burial site in Črnomelj was active between the 13th and 18th century, whereas the more recent Polje burial was in use from the 16th to 19th century, creating different temporal and geographical environments. Results for the Črnomelj burial site revealed that the petrous bone outperformed all other bone types studied, except the calcaneus. At the Polje archeological site calcanei, tali, and femurs yielded the same STR typing success as petrous bones. The results obtained highlight the importance of careful bone sample selection for DNA analysis of aged skeletal remains. In addition to petrous bones, calcanei were found to be an alternative source of DNA when older burial sites are investigated. When more recent burial sites are processed, calcanei, tali, and femurs should be sampled besides petrous bones, not only because they exhibited good performance, but also because of easier sampling and easier grinding in the case of trabecular bones. This study contributes valuable insights into the potential use of various skeletal types as a source of DNA for investigation of aged skeletal remains, and it offers practical implications for forensic and archaeological investigations.
岩骨比人体其他任何骨骼都含有更高浓度的 DNA。由于采样具有很强的破坏性，而且并非总是能获得完整的遗骸，因此需要寻找其他来源的 DNA。为了确定其他最佳的骨骼类型，我们比较了从两个不同现代时代基督教墓地的 66 具成人骨骼中采集的岩骨、股骨、距骨和跟骨。采用全脱矿化提取方法获取 DNA，实时 PCR 定量法确定 DNA 量和降解程度，以及商业化法医短串联重复序列（STR）PCR 扩增试剂盒确定遗传图谱。通过统计分析来探讨 DNA 产量、DNA 降解和 STR 扩增成功率方面的差异。虽然很少有研究对骨骼内 DNA 保存的变异性进行系统探索，包括不同的挖掘地点因时间和地理位置而异，但调查的第二部分是基于对两个考古地点的比较，这使我们能够比较不同的死后间隔和环境条件对 DNA 保存的影响。Črnomelj 较古老的墓地活跃于 13 至 18 世纪，而较新的 Polje 墓地使用于 16 至 19 世纪，创造了不同的时间和地理环境。来自Črnomelj 墓地的研究结果表明，岩骨表现优于除跟骨以外的所有其他研究骨骼类型。在 Polje 考古遗址中，跟骨、距骨和股骨与岩骨具有相同的 STR 分型成功率。研究结果强调了在对老年骨骼遗骸进行 DNA 分析时，需要仔细选择骨骼样本。除岩骨外，当研究较古老的墓地时，跟骨也可作为 DNA 的替代来源。当处理较新的墓地时，除岩骨外，还应采集跟骨、距骨和股骨，不仅因为它们表现良好，而且因为在涉及松质骨时，这些骨骼更容易采样和研磨。这项研究为各种骨骼类型作为老年骨骼遗骸研究的 DNA 来源的潜在用途提供了有价值的见解，并为法医和考古调查提供了实际意义。
