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应用 RNAscope 技术研究中国猪流行性腹泻病毒变异株 BJ2011C 在新生仔猪中的感染动态。

Application of RNAscope technology to studying the infection dynamics of a Chinese porcine epidemic diarrhea virus variant strain BJ2011C in neonatal piglets.

机构信息

Key Laboratory of Animal Epidemiology of the Ministry of Agriculture, College of Veterinary Medicine, China Agricultural University, Beijing 100193, People's Republic of China.

Key Laboratory of Animal Epidemiology of the Ministry of Agriculture, College of Veterinary Medicine, China Agricultural University, Beijing 100193, People's Republic of China.

出版信息

Vet Microbiol. 2019 Aug;235:220-228. doi: 10.1016/j.vetmic.2019.07.003. Epub 2019 Jul 8.

Abstract

The highly virulent porcine epidemic diarrhea virus (PEDV) variants cause the death of mainly neonatal piglets, but how the viruses spread within the gastro-intestinal tract in a temporal and spatial manner has remained poorly characterized but is critical to understand the viral pathogenesis. In this study, we used the Chinese PEDV epidemic strain BJ2011C as a model organism and took advantage of the newly developed RNAscope in situ hybridization technology to investigate the tempo-spatial infection dynamics in neonatal piglets. We found that the PEDV strain BJ2011C could quickly colonize the small intestine, which occurred in just 6 h post infection, with virus shedding starting at 6 hpi and peaking at 24 hpi. Jejunum was the first target tissue for infection and then ileum, followed by infrequent infection of duodenum. In these tissues, the virus nucleic acids were mainly present in the villous epithelial cells but not in crypt cells. Interestingly, the viral RNAs were not detectable by RNAscope in large intestines although tissue damages could be discerned by H & E staining. Overall, our results provide useful information about spread dynamics and tissue preference of PEDV epidemic strain BJ2011C.

摘要

高致病性猪流行性腹泻病毒(PEDV)变异株主要导致仔猪死亡,但病毒如何在胃肠道中随时间和空间传播仍知之甚少,但这对于了解病毒发病机制至关重要。在本研究中,我们使用中国 PEDV 流行株 BJ2011C 作为模型生物,并利用新开发的 RNAscope 原位杂交技术来研究新生仔猪的时空感染动态。我们发现 PEDV 株 BJ2011C 可以快速定植小肠,在感染后 6 小时即可发生,病毒脱落从 6 小时开始,24 小时达到高峰。空肠是感染的第一靶组织,然后是回肠,接着是十二指肠的偶发感染。在这些组织中,病毒核酸主要存在于绒毛状上皮细胞中,而不是隐窝细胞中。有趣的是,尽管可以通过 H & E 染色识别组织损伤,但 RNAscope 无法在大肠中检测到病毒 RNA。总的来说,我们的结果提供了有关 PEDV 流行株 BJ2011C 传播动态和组织偏好的有用信息。

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