Zurich Institute of Forensic Medicine, University of Zurich, Switzerland.
Institute of Legal Medicine, University Hospital, University of Ulm, Germany.
Forensic Sci Int Genet. 2021 Jan;50:102409. doi: 10.1016/j.fsigen.2020.102409. Epub 2020 Oct 26.
In recent years, forensic mRNA profiling has increasingly been used to identify the origin of human body fluids. By now, several laboratories have implemented mRNA profiling and also use it in criminal casework. In 2018 the FoRNAP (Forensic RNA Profiling) group was established among a number of these laboratories with the aim of sharing experiences, discussing optimization potential, identifying challenges and suggesting solutions with regards to mRNA profiling and casework. To compare mRNA profiling methods and results a collaborative exercise was organized within the FoRNAP group. Seven laboratories from four countries received 16 stains, comprising six pure body fluid / tissue stains and ten mock casework samples. The laboratories were asked to analyze the provided stains with their in-house method (PCR/CE or MPS) and markers of choice. Five laboratories used a DNA/RNA co-extraction strategy. Overall, up to 11 mRNA markers per body fluid were analyzed. We found that mRNA profiling using different extraction and analysis methods as well as different multiplexes can be applied to casework-like samples. In general, high input samples were typed with high accuracy by all laboratories, regardless of the method used. Irrespective of the analysis strategy, samples of low input or mixed stains were more challenging to analyze and interpret since, alike to DNA profiling, a higher number of markers dropped out and/or additional unexpected markers not consistent with the cell type in question were detected. It could be shown that a plethora of different but valid analysis and interpretation strategies exist and are successfully applied in the Forensic Genetics community. Nevertheless, efforts aiming at optimizing and harmonizing interpretation approaches in order to achieve a higher consistency between laboratories might be desirable in the future. The simultaneous extraction of DNA alongside RNA showed to be an effective approach to identify not only the body fluid present but also to identify the donor(s) of the stain. This allows investigators to gain valuable information about the origin of crime scene samples and the course of events in a crime case.
近年来,法医信使 RNA 分析越来越多地被用于鉴定人体体液的来源。目前,已有几家实验室实施了信使 RNA 分析,并将其应用于刑事案件中。2018 年,一些实验室成立了 FoRNAP(法医信使 RNA 分析)小组,旨在分享经验,讨论优化潜力,识别挑战,并就信使 RNA 分析和案例工作提出解决方案。为了比较信使 RNA 分析方法和结果,FoRNAP 小组组织了一次协作活动。来自四个国家的七个实验室收到了 16 个样本,包括六种纯体液/组织样本和十种模拟案例样本。实验室被要求使用内部方法(PCR/CE 或 MPS)和选择的标记物分析提供的样本。五个实验室使用了 DNA/RNA 联合提取策略。总体而言,每个体液分析了多达 11 个信使 RNA 标记物。我们发现,使用不同的提取和分析方法以及不同的多重标记物的信使 RNA 分析可以应用于类似案例的样本。通常,所有实验室都能以高精度对高输入样本进行分型,无论使用何种方法。无论分析策略如何,低输入或混合样本更具挑战性,因为与 DNA 分析一样,更多的标记物丢失,并且/或者检测到与所涉及的细胞类型不一致的额外意外标记物。可以证明,存在大量不同但有效的分析和解释策略,并且在法医遗传学领域成功应用。然而,未来可能需要努力优化和协调解释方法,以提高实验室之间的一致性。同时提取 DNA 和 RNA 是一种有效的方法,不仅可以识别存在的体液,还可以识别样本的供体。这使调查人员能够获得有关犯罪现场样本来源和犯罪案件中事件进程的有价值信息。