Song Zhenhui, Dai Xianjin, Ye Cuifang, Li Yuntian, Wang Li, Hu Yang
Department of Veterinary Medicine, Southwest University, Rongchang Campus, Chongqing 402460, PR China.
J Virol Methods. 2016 Dec;238:6-12. doi: 10.1016/j.jviromet.2016.09.018. Epub 2016 Sep 28.
To gain a better understanding of the replication, proliferation and infection characteristics of porcine transmissible gastroenteritis virus (TGEV) in porcine intestinal epithelial cells (IECs), this study established a cell model of IECs infected with the Chongqing (CQ) strain of TGEV. The morphogenesis and proliferative rule of TGEV in porcine IECs were investigated using transmission electron microscopy, indirect immunofluorescence assays and real-time fluorescence quantitative PCR. Observations under the TEM indicated that the enveloped viral particles were roughly spherical, with diameters of between 80 and 120nm. The virions entered porcine IECs by membrane fusion and the mature viruses in the vacuoles were transported to the cell membrane before release. The results also showed that from 0 to 12h after TGEV infection of porcine IECs, the intracellular viral RNA content did not change significantly. Logarithmic growth occurred from 12 to 36h, after which it gradually decreased. Moreover, the extracellular RNA content began to rise at 24h after inoculation and then reduced gradually at approximately 48h. This study provided a theoretical foundation for further study on the infection characteristics of TGEV in target cells.
为了更好地了解猪传染性胃肠炎病毒(TGEV)在猪肠道上皮细胞(IECs)中的复制、增殖和感染特性,本研究建立了感染TGEV重庆(CQ)株的IECs细胞模型。利用透射电子显微镜、间接免疫荧光试验和实时荧光定量PCR研究了TGEV在猪IECs中的形态发生和增殖规律。透射电镜观察表明,包膜病毒粒子大致呈球形,直径在80至120nm之间。病毒粒子通过膜融合进入猪IECs,空泡中的成熟病毒在释放前被转运到细胞膜。结果还表明,TGEV感染猪IECs后0至12小时,细胞内病毒RNA含量无明显变化。12至36小时出现对数增长,之后逐渐下降。此外,接种后24小时细胞外RNA含量开始上升,然后在约48小时逐渐下降。本研究为进一步研究TGEV在靶细胞中的感染特性提供了理论基础。