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Long-term room temperature preservation of corpse soft tissue: an approach for tissue sample storage

机译：尸体软组织的长期室温保存：组织样本存储的一种方法

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Background Disaster victim identification (DVI) represents one of the most difficult challenges in forensic sciences, and subsequent DNA typing is essential. Collected samples for DNA-based human identification are usually stored at low temperature to halt the degradation processes of human remains. We have developed a simple and reliable procedure for soft tissue storage and preservation for DNA extraction. It ensures high quality DNA suitable for PCR-based DNA typing after at least 1 year of room temperature storage. Methods Fragments of human psoas muscle were exposed to three different environmental conditions for diverse time periods at room temperature. Storage conditions included: (a) a preserving medium consisting of solid sodium chloride (salt), (b) no additional substances and (c) garden soil. DNA was extracted with proteinase K/SDS followed by organic solvent treatment and concentration by centrifugal filter devices. Quantification was carried out by real-time PCR using commercial kits. Short tandem repeat (STR) typing profiles were analysed with 'expert software'. Results DNA quantities recovered from samples stored in salt were similar up to the complete storage time and underscored the effectiveness of the preservation method. It was possible to reliably and accurately type different genetic systems including autosomal STRs and mitochondrial and Y-chromosome haplogroups. Autosomal STR typing quality was evaluated by expert software, denoting high quality profiles from DNA samples obtained from corpse tissue stored in salt for up to 365 days. Conclusions The procedure proposed herein is a cost efficient alternative for storage of human remains in challenging environmental areas, such as mass disaster locations, mass graves and exhumations. This technique should be considered as an additional method for sample storage when preservation of DNA integrity is required for PCR-based DNA typing.
机译：背景技术灾难受害者识别（DVI）是法医学领域最困难的挑战之一，随后的DNA分型至关重要。通常将收集的样本用于基于DNA的人类识别，然后将其低温保存，以阻止人类残骸的降解过程。我们已经开发出一种简单而可靠的程序，用于软组织的存储和DNA提取的保存。室温保存至少一年后，可确保高质量的DNA适合基于PCR的DNA分型。方法在室温下，将人腰肌的碎片暴露于三种不同的环境条件下不同的时间段。储存条件包括：（a）由固体氯化钠（盐）组成的保存介质，（b）无其他物质，以及（c）园林土壤。用蛋白酶K / SDS提取DNA，然后进行有机溶剂处理并通过离心过滤器进行浓缩。使用商业试剂盒通过实时PCR进行定量。使用“专家软件”分析了短串联重复序列（STR）的基因型。结果从盐中存储的样品中回收的DNA量在整个存储时间内都相似，这突出了保存方法的有效性。可以可靠，准确地键入不同的遗传系统，包括常染色体STR，线粒体和Y染色体单倍型。通过专家软件评估常染色体STR分型质量，这表明从盐中存储的尸体组织中提取的DNA样品（长达365天）的高质量概况。结论本文提出的程序是在具有挑战性的环境区域（例如大规模灾难地点，大规模坟墓和尸体发掘）中存储人类遗体的一种经济高效的替代方法。当基于PCR的DNA分型需要保持DNA完整性时，该技术应被视为样品存储的另一种方法。

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