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首页> 外文期刊>Investigative Genetics >Targeted sampling of cementum for recovery of nuclear DNA from human teeth and the impact of common decontamination measures
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Targeted sampling of cementum for recovery of nuclear DNA from human teeth and the impact of common decontamination measures

机译：有针对性的牙骨质采样以从人的牙齿中回收核DNA以及常规去污措施的影响

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Background Teeth are a valuable source of DNA for identification of fragmented and degraded human remains. While the value of dental pulp as a source of DNA is well established, the quantity and presentation of DNA in the hard dental tissues has not been extensively studied. Without this knowledge common decontamination, sampling and DNA extraction techniques may be suboptimal. Targeted sampling of specific dental tissues could maximise DNA profiling success, while minimising the need for laborious sampling protocols and DNA extraction techniques, thus improving workflows and efficiencies. We aimed to determine the location of cellular DNA in non-degraded human teeth to quantify the yield of nuclear DNA from cementum, the most accessible and easily sampled dental tissue, and to investigate the effect of a common decontamination method, treatment with sodium hypochlorite (bleach). We examined teeth histologically and subsequently quantified the yield of nuclear DNA from the cementum of 66 human third molar teeth. We also explored the effects of bleach (at varying concentrations and exposure times) on nuclear DNA within teeth, using histological and quantitative PCR methods. Results Histology confirmed the presence of nucleated cells within pulp and cementum, but not in dentine. Nuclear DNA yields from cementum varied substantially between individuals but all samples gave sufficient DNA (from as little as 20 mg of tissue) to produce full short tandem repeat (STR) profiles. Variation in yield between individuals was not influenced by chronological age or sex of the donor. Bleach treatment with solutions as dilute as 2.5% for as little as 1 min damaged the visible nuclear material and reduced DNA yields from cementum by an order of magnitude. Conclusions Cementum is a valuable, and easily accessible, source of nuclear DNA from teeth, and may be a preferred source where large numbers of individuals need to be sampled quickly (for example, mass disaster victim identification) without the need for specialist equipment or from diseased and degraded teeth, where pulp is absent. Indiscriminant sampling and decontamination protocols applied to the outer surface of teeth can destroy this DNA, reducing the likelihood of successful STR typing results.
机译：背景技术牙齿是DNA的宝贵来源，可用于鉴定碎片化和降解的人类遗骸。虽然牙髓作为DNA来源的价值已得到公认，但尚未对硬牙组织中DNA的数量和呈递进行广泛研究。没有这种知识，常见的去污，采样和DNA提取技术可能不够理想。特定牙齿组织的目标采样可以最大程度地提高DNA谱图的成功率，同时最大程度地减少对费力的采样协议和DNA提取技术的需求，从而改善工作流程和效率。我们的目的是确定细胞DNA在未降解的人类牙齿中的位置，以量化牙骨质（最易接近和易于采样的牙组织）中核DNA的产量，并研究使用次氯酸钠处理普通去污方法的效果（漂白）。我们通过组织学检查了牙齿，随后定量了66颗人类第三磨牙的牙骨质的核DNA产量。我们还使用组织学和定量PCR方法探索了漂白剂（浓度和暴露时间不同）对牙齿核DNA的影响。结果组织学证实牙髓和牙骨质中存在有核细胞，但牙本质中不存在。牙骨质的核DNA产量在个体之间差异很大，但所有样品均提供足够的DNA（低至20 mg组织）以产生完整的短串联重复序列（STR）。个体之间产量的变化不受捐助者的年龄或性别的影响。用稀释至2.5％的溶液漂白处理至少1分钟会破坏可见的核材料，并使牙骨质的DNA产量降低一个数量级。结论牙骨质是牙齿中有价值的，易于获取的核DNA来源，并且可能是首选的来源，无需大量专业设备或其他工具就可以快速采样大量个体（例如，大规模灾难受害者识别）。患病和退化的牙齿，缺牙髓。应用于牙齿外表面的不加区别的采样和去污协议会破坏该DNA，从而降低成功进行STR分型结果的可能性。

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