Department of Forensic Genetics, West China School of Basic Medical Sciences & Forensic Medicine, Sichuan University, Chengdu, 610041, Sichuan, People's Republic of China.
Laboratory of Molecular Translational Medicine, Center for Translational Medicine, Key Laboratory of Birth Defects and Related Diseases of Women and Children (Sichuan University), Ministry of Education, West China Second University Hospital, Sichuan University, Chengdu, Sichuan, 610041, People's Republic of China.
Int J Legal Med. 2022 Nov;136(6):1565-1575. doi: 10.1007/s00414-022-02886-y. Epub 2022 Sep 8.
Analysis of genetic markers can provide clues for case investigation. Short tandem repeat (STR) detection and analysis are widely used for both personal identification and parentage testing. However, DNA analysis currently cannot provide sufficient information for body fluid identification. Tissue or cell sources of samples can be identified by detecting body fluid-specific mRNA markers, which have been studied thoroughly. Integrating STR profiling and mRNA expression patterns can provide more information than conventional methods for investigations and the reconstruction of crime scenes; this can be achieved by DNA/RNA co-extraction technology, which is economical, efficient, and suitable for low-template samples. Here, we propose a co-analysis system based on the PowerPlex 16 kit. This system can simultaneously amplify 25 markers, including 15 STRs, one non-STR amelogenin, and nine mRNA markers (three blood-specific, two saliva-specific, two semen-specific, and two housekeeping gene markers). The specificity and sensitivity of the co-analysis system were determined and aged and degraded samples were used to validate the stability of the co-analysis system. Finally, different DNA/RNA ratios and various carriers were evaluated. The results showed that the DNA/RNA co-analysis system correctly identified different types of body fluid stains. The STR profiles obtained using the co-analysis system were identical to those obtained using the PP16 kit, which demonstrates that the mRNA primers used did not affect STR profiling. Complete STR and mRNA profiles could be obtained from 1/8 portions of buccal swabs, 1/16 portions of swabs of blood and semen samples, 0.1 cm of blood samples, 0.25 cm of semen samples, and 1.0 cm saliva samples. Additionally, our findings indicate that complete STR and mRNA profiles can be obtained with this system from blood and semen samples when the DNA/RNA ratio is 1:1/32. This study suggests that the co-analysis system could be used for simultaneous personal identification and body fluid identification.
遗传标记分析可为案件调查提供线索。短串联重复序列(STR)检测和分析广泛应用于个人识别和亲子关系测试。然而,目前的 DNA 分析无法为体液鉴定提供足够的信息。可以通过检测体液特异性 mRNA 标记物来鉴定组织或细胞来源的样本,这些标记物已经得到了深入研究。整合 STR 谱分析和 mRNA 表达模式可以为调查和犯罪现场重建提供比传统方法更多的信息;这可以通过经济高效且适合低模板样本的 DNA/RNA 共提取技术来实现。在这里,我们提出了一种基于 PowerPlex 16 试剂盒的共分析系统。该系统可以同时扩增 25 个标记物,包括 15 个 STR、一个非 STR 牙本质蛋白和 9 个 mRNA 标记物(3 个血液特异性、2 个唾液特异性、2 个精液特异性和 2 个管家基因标记物)。我们确定了共分析系统的特异性和灵敏度,并使用老化和降解的样本验证了共分析系统的稳定性。最后,我们评估了不同的 DNA/RNA 比值和各种载体。结果表明,该 DNA/RNA 共分析系统能够正确识别不同类型的体液斑。使用共分析系统获得的 STR 谱与使用 PP16 试剂盒获得的 STR 谱完全一致,这表明使用的 mRNA 引物不会影响 STR 谱分析。从 1/8 份口腔拭子、1/16 份血液和精液样本拭子、0.1 cm 血液样本、0.25 cm 精液样本和 1.0 cm 唾液样本中可以获得完整的 STR 和 mRNA 谱。此外,我们的研究结果表明,当 DNA/RNA 比值为 1:1/32 时,该系统可以从血液和精液样本中获得完整的 STR 和 mRNA 谱。本研究表明,该共分析系统可用于同时进行个人识别和体液识别。