Department of Anthropology, 1621 Cumberland Avenue, 502A Strong Hall, University of Tennessee, Knoxville, TN, 37996-1525, United States.
International Commission on Missing Persons Headquarters, Koninginnegracht 12 2514 AA Den Haag, The Netherlands; Tyrolpath, Klostergasse 1, 6511 Zams, Austria.
Forensic Sci Int Genet. 2018 Sep;36:167-175. doi: 10.1016/j.fsigen.2018.07.009. Epub 2018 Jul 10.
In mass fatality events, the need to identify large numbers of deceased persons using DNA can be a significant drain on already overburdened forensic practitioners, both in the field setting and the laboratory. The laboratory may be required to extract DNA from a variety of postmortem sample types, family or direct reference samples related to the missing, and perform matching of these results in a short period of time. While most forensic institutions are well equipped to handle both family and direct reference samples, postmortem samples such as bone or heterogeneous tissue samples can be difficult for labs to analyze. We have devised an easily deployable, efficient, and inexpensive method for collecting postmortem DNA samples on commercially available DNA preservation cards ("FTA" cards). FTA cards are already widely used in forensic labs and are convenient for shipping due to their small volume and stability at room temperature. We evaluated the suitability of a protocol involving swabbing of incisions made on cadavers and sample deposition onto FTA cards over various postmortem intervals and under different environmental conditions. Each trial took place during a different point in the calendar year to evaluate the effects of seasonal weather patterns and temperature on decomposition, DNA yield, and rates of DNA degradation. To further account for the effects of seasonality (temperature and humidity), the progression of body decomposition was recorded following the Total Body Score (TBS) method [1]. DNA degradation was assessed either through STR amplification of 1.2 mm FTA punches or DNA extraction from 3.0 mm punches followed by real-time PCR quantification and STR amplification and genotyping. No consistent relationship was observed between postmortem interval and DNA degradation. Instead, the TBS score, which captures the stage of body decomposition, was shown to correlate well with DNA quantity. A TBS of 15 and below consistently yielded strong partial or full profiles (20 STR loci and Amelogenin using the PowerPlex 21 System) from all individuals from either 1.2 mm or 3.0 mm punches. Transfer of sample swabs to FTA cards is shown to be a simple and effective method for both field and laboratory operations over a range of conditions that can be evaluated by field forensic practitioners based on a body decomposition score. The approach could be beneficially integrated into mass fatality response plans.
在大规模死亡事件中,需要使用 DNA 识别大量死者,这可能会给已经负担过重的法医从业人员带来巨大压力,无论是在现场环境还是实验室中。实验室可能需要从各种死后样本类型、与失踪人员有关的家庭或直接参考样本中提取 DNA,并在短时间内进行这些结果的匹配。虽然大多数法医机构都有能力处理家庭和直接参考样本,但骨骼或异质组织等死后样本可能难以分析。我们设计了一种简单、高效且经济的方法,用于在市售的 DNA 保存卡(“FTA”卡)上收集死后 DNA 样本。FTA 卡已经在法医实验室中广泛使用,由于其体积小且在室温下稳定,因此便于运输。我们评估了一种方案的适用性,该方案涉及在死后不同时间间隔和不同环境条件下,对尸体切口进行拭子取样,并将样本沉积到 FTA 卡上。每次试验都在日历年度的不同时间点进行,以评估季节性天气模式和温度对分解、DNA 产量和 DNA 降解率的影响。为了进一步考虑季节性(温度和湿度)的影响,按照全身评分(TBS)方法[1]记录了尸体分解的进展情况。通过对 1.2mm FTA 打孔的 STR 扩增或对 3.0mm 打孔的 DNA 提取进行实时 PCR 定量和 STR 扩增和基因分型来评估 DNA 降解。没有观察到死后间隔时间和 DNA 降解之间存在一致的关系。相反,TBS 评分(捕捉身体分解的阶段)与 DNA 数量密切相关。TBS 评分在 15 及以下时,从所有个体的 1.2mm 或 3.0mm 打孔中均能获得强的部分或完整的图谱(使用 PowerPlex 21 系统的 20 个 STR 基因座和 amelogenin)。从现场法医从业人员的角度来看,将样本拭子转移到 FTA 卡上是一种简单有效的方法,可以在一系列条件下进行评估。该方法可有效地整合到大规模死亡事件应对计划中。