Division of Viral Disease and Epidemiology, National Institute of Animal Health, National Agriculture and Food Research Organization, 3-1-5 Kannondai, Tsukuba, Ibaraki, 305-0856, Japan.
Division of Viral Disease and Epidemiology, National Institute of Animal Health, National Agriculture and Food Research Organization, 3-1-5 Kannondai, Tsukuba, Ibaraki, 305-0856, Japan.
J Virol Methods. 2021 Apr;290:114068. doi: 10.1016/j.jviromet.2021.114068. Epub 2021 Jan 15.
Porcine deltacoronavirus (PDCoV) is a novel swine enteropathogenic coronavirus that leads to acute diarrhea/vomiting, dehydration, and mortality in seronegative neonatal piglets. As widely known, attempts to culture porcine enteropathogenic coronaviruses, such as PDCoV and porcine epidemic diarrhea virus, in cells have been proven to be difficult. This study aimed to establish an efficient and cost-effective culture system for PDCoV using embryonated chicken eggs (ECEs) to enable future vaccine production and efficient virus isolation from infected animals. The inoculation of samples into the allantoic cavity of 3- to 7-day-old ECEs yielded efficient virus propagation even from porcine fecal samples. Virus propagation in 2- and 8-day-old ECEs were confirmed in 30.0 % and 11.1 % of the samples, respectively. This indicates that susceptible cells rapidly develop in 2-day-old ECEs and differentiate to mature cells that are nonsusceptible to PDCoV in 8-day-old layer chicken ECEs. Furthermore, our study demonstrated that PDCoV can be passaged in 6-day-old ECEs with high viral replicative efficiency. This technique for propagating PDCoV using ECEs is a powerful tool that could be utilized for PDCoV vaccine development and virus isolation from poultry, livestock, and wild animals.
猪德尔塔冠状病毒(PDCoV)是一种新型的猪肠道致病性冠状病毒,可导致血清阴性的新生仔猪发生急性腹泻/呕吐、脱水和死亡。众所周知,尝试在细胞中培养猪肠道致病性冠状病毒,如 PDCoV 和猪流行性腹泻病毒,已被证明是困难的。本研究旨在建立一种使用鸡胚(ECE)的高效且经济有效的 PDCoV 培养系统,以实现未来的疫苗生产和从感染动物中有效分离病毒。将样品接种到 3-7 日龄鸡胚的尿囊腔中,即使来自猪粪便样本,也能有效繁殖病毒。在 2 日龄和 8 日龄鸡胚中分别确认了 30.0%和 11.1%的样品中病毒的繁殖。这表明易感细胞在 2 日龄鸡胚中迅速发育,并在 8 日龄蛋鸡鸡胚中分化为对 PDCoV 不敏感的成熟细胞。此外,我们的研究表明,PDCoV 可以在 6 日龄鸡胚中进行传代,具有很高的病毒复制效率。使用鸡胚繁殖 PDCoV 的这种技术是一种强大的工具,可用于 PDCoV 疫苗开发和从家禽、牲畜和野生动物中分离病毒。