Department of Diagnostics and Public Health, Legal Medicine Section, University of Verona, 37134 Verona, Italy.
Department of Diagnostics and Public Health, Microbiology Section, University of Verona, 37134 Verona, Italy.
Front Biosci (Landmark Ed). 2022 Jun 7;27(6):183. doi: 10.31083/j.fbl2706183.
In many forensic cases, the medical records of the deceased are not available at the time of the autopsy; therefore, no information about the deceased's state of health, including any infectious diseases contracted during life, is accessible. The detection of some of the principal viral infections, such as hepatitis B virus (HBV), hepatitis C virus (HCV), and human immunodeficiency virus type 1 (HIV-1), could contribute to determining causes of death and interesting applications could be found in medico-legal practice, such as occupational risk assessment. To date, accurate and sensitive serological and molecular assays capable of detecting these viruses have been validated on biological samples taken from living beings, while their efficiency on forensic post-mortem biological samples has yet to be thoroughly assessed. To further this aim, this study evaluated whether the nucleic acid amplification techniques (NAATs) for the detection of viral genomes that are applied in clinical settings can be used, with the same success rate, for these latter samples.
Manual viral nucleic acid extraction processes and fully-automated amplification-based detection techniques developed in-house were evaluated on blood samples taken during the routine autopsies of 21 cadavers performed 2 to 9 days after death. Information on HBV, HCV, and HIV-1 seropositive status was previously known for only four of these cadavers.
Using automated quantitative real-time PCR (qPCR) and qualitative PCR (end-point) analyses, it was possible to confirm the presence of viral genomes in the four post-mortem whole blood samples with previously reported specific serological positivity. In addition, the genomes of HCV and/or HIV-1 genomes were detected in three other blood samples with unknown serological status at the time of autopsy.
Therefore, our findings suggest that molecular assays may detect the presence of viral genomes in forensic post-mortem blood samples up to five days after death. This provides an additional means of investigation that can contribute to the determination of the deceased's cause of death.
在许多法医学案例中，尸检时无法获得死者的病历；因此，无法获取死者的健康状况信息，包括生前患有的任何传染病。检测一些主要的病毒感染，如乙型肝炎病毒（HBV）、丙型肝炎病毒（HCV）和人类免疫缺陷病毒 1 型（HIV-1），可以帮助确定死因，并在医学法律实践中发现有趣的应用，如职业风险评估。迄今为止，已经在取自活体的生物样本上验证了准确且敏感的可检测这些病毒的血清学和分子检测方法，但其在法医死后生物样本中的效率尚未得到彻底评估。为了进一步实现这一目标，本研究评估了用于检测病毒基因组的核酸扩增技术（NAATs）是否可以以相同的成功率应用于这些后者样本。
手动病毒核酸提取过程和内部开发的全自动化扩增检测技术在 21 例常规尸检中进行了评估，这些尸检是在死后 2 至 9 天进行的。这些尸体中只有 4 例之前已知 HBV、HCV 和 HIV-1 血清阳性状态。
使用自动化定量实时 PCR（qPCR）和定性 PCR（终点）分析，能够在先前报告具有特定血清学阳性的 4 例死后全血样本中确认病毒基因组的存在。此外，在另外 3 例尸检时未知血清学状态的血液样本中检测到 HCV 和/或 HIV-1 基因组。
因此，我们的发现表明，分子检测方法可以在死后五天内检测到法医死后血液样本中病毒基因组的存在。这提供了一种额外的调查手段，可以帮助确定死者的死因。
