Louisiana State University Agricultural Center, Agricultural Experiment Station, Department of Entomology, 415 Life Sciences, Baton Rouge, Louisiana 70803, USA.
Louisiana State University Agricultural Center, Agricultural Experiment Station, Bob R. Jones Idlewild Research Station, 4419 Idlewild Drive, Clinton, Louisiana 70722, USA.
J Wildl Dis. 2020 Jan;56(1):58-65. Epub 2019 Aug 12.
We determined the temporal aspects of detecting bluetongue virus (BTV) and epizootic hemorrhagic disease virus (EHDV) in postmortem bone marrow samples of white-tailed deer () using molecular and in vitro cell culture techniques. Bone marrow samples from carcasses were collected and assayed on the day of death and at intervals up to 16 wk after death. We recovered BTV and EHDV from fresh bone marrow collected at day 0 by isolation in Vero and BHK-21 cell cultures. However, attempts to replicate the viruses from aged bone marrow in Vero and BHK-21 cell cultures failed. The real-time quantitative reverse transcriptase PCR (qRT-PCR) results confirmed that EHDV and BTV can be detected in aged bone marrow for up to 12 and 16 wk, respectively, after death. The RNA of BTV and EHDV could be detected by qRT-PCR in white-tailed deer bone marrow for extended periods of time postmortem. This technique will provide a useful tool for retrospective determination of BTV or EHDV infection of white-tailed deer at the time of death.
我们使用分子和体外细胞培养技术，确定了死后骨髓样本中蓝舌病病毒（BTV）和流行性出血热病毒（EHDV）的时间特征。从尸体采集骨髓样本，并在死亡当天以及死亡后长达 16 周的时间内进行检测。我们从 0 天新鲜采集的骨髓中通过在 Vero 和 BHK-21 细胞培养物中分离，成功分离到了 BTV 和 EHDV。然而，从老化的骨髓中在 Vero 和 BHK-21 细胞培养物中复制病毒的尝试均失败了。实时定量逆转录聚合酶链反应（qRT-PCR）结果证实，EHDV 和 BTV 分别可在死后长达 12 周和 16 周的时间内，从老化的骨髓中检测到。BTV 和 EHDV 的 RNA 可通过 qRT-PCR 在白尾鹿死后的很长一段时间的骨髓中检测到。该技术将为追溯性确定白尾鹿死亡时的 BTV 或 EHDV 感染提供有用的工具。
