Erich and Hanna Klessmann-Institute for Cardiovascular Research and Development, Heart- and Diabetes Center NRW, University Hospital of the Ruhr-University Bochum, Georgstr. 11, D-32545, Bad Oeynhausen, Germany.
Institute for Anesthesiology, Intensive Care- and Emergency Medicine, Johannes Wesling Hospital Minden, MKK-Hospital, Campus OWL, Ruhr-University Bochum, Bochum, Germany.
Int J Legal Med. 2023 Sep;137(5):1569-1581. doi: 10.1007/s00414-023-02966-7. Epub 2023 Feb 11.
The out-of-hospital cardiac arrest (OHCA) in the young may be associated with a genetic predisposition which is relevant even for genetic counseling of relatives. The identification of genetic variants depends on the availability of intact genomic DNA. DNA from autopsy may be not available due to low autopsy frequencies or not suitable for high-throughput DNA sequencing (NGS). The emergency medical service (EMS) plays an important role to save biomaterial for subsequent molecular autopsy. It is not known whether the DNA integrity of samples collected by the EMS is better suited for NGS than autopsy specimens.
DNA integrity was analyzed by standardized protocols. Fourteen blood samples collected by the EMS and biomaterials from autopsy were compared. We collected 172 autopsy samples from different tissues and blood with postmortem intervals of 14-168 h. For comparison, DNA integrity derived from blood stored under experimental conditions was checked against autopsy blood after different time intervals.
DNA integrity and extraction yield were higher in EMS blood compared to any autopsy tissue. DNA stability in autopsy specimens was highly variable and had unpredictable quality. In contrast, collecting blood samples by the EMS is feasible and delivered comparably the highest DNA integrity.
Isolation yield and DNA integrity from blood samples collected by the EMS is superior in comparison to autopsy specimens. DNA from blood samples collected by the EMS on scene is stable at room temperature or even for days at 4 °C. We conclude that the EMS personnel should always save a blood sample of young fatal OHCA cases died on scene to enable subsequent genetic analysis.
院外心脏骤停(OHCA)在年轻人中可能与遗传易感性有关,即使对于亲属的遗传咨询也是如此。遗传变异的识别取决于完整基因组 DNA 的可用性。由于尸检频率低或不适合高通量 DNA 测序(NGS),可能无法获得尸检的 DNA。紧急医疗服务(EMS)在保存生物材料以进行后续分子尸检方面发挥着重要作用。尚不清楚 EMS 采集的样本的 DNA 完整性是否比尸检标本更适合 NGS。
通过标准化方案分析 DNA 完整性。比较了 EMS 采集的 14 个血液样本和尸检生物材料。我们从不同组织和血液中收集了 172 个尸检样本,死后间隔时间为 14-168 小时。为了比较,检查了在不同时间间隔后从实验条件下储存的血液中得出的 DNA 完整性与尸检血液中的 DNA 完整性。
与任何尸检组织相比,EMS 血液中的 DNA 完整性和提取产量更高。尸检标本中的 DNA 稳定性差异很大,质量不可预测。相比之下,通过 EMS 收集血液样本是可行的,并提供了相当高的 DNA 完整性。
与尸检标本相比,EMS 采集的血液样本的分离产量和 DNA 完整性更高。EMS 现场采集的血液样本在室温下稳定,甚至在 4°C 下数天也稳定。我们得出的结论是,EMS 人员应始终保存现场死亡的年轻致命 OHCA 病例的血液样本,以进行后续的遗传分析。