猪流行性腹泻病毒SHpd/2012株侵染Vero细胞特性分析
Characteristics of vero cells infected by SHpd/2012 strain of porcine epidemic diarrhea virus

为了探究PEDV(猪流行性腹泻病毒)流行毒株SHpd/2012侵染细胞的特性,研究采用IFA(间接免疫荧光)及绘制病毒一步生长曲线等方法,对感染PEDV SHpd/2012株后的Vero细胞(非洲绿猴肾细胞)病变过程进行研究。结果发现,CPE(细胞病变)主要表现为细胞肿胀、变圆、皱缩、聚集成合胞体甚至从培养瓶壁大量脱落。IFA结果显示,接毒后12 h病毒开始增殖,在接毒后36 h时感染细胞数量最多,其后病毒增殖速度逐步下降,60 h时趋于平稳。病毒一步生长曲线结果显示,SHpd/2012株感染细胞后24~60 h的毒价一直处于较高水平,60 h后开始下降。此外,本研究成功克隆并真核表达了PEDV S蛋白,为研究PEDV与受体蛋白的互作及病毒的致病机制奠定基础。
IFA(immunoflorescent assay)and growth curves was employed to study the characteristics of Vero cells infected by SHpd/2012 strain of PEDV(porcine epidemic diarrhea virus).The infected cells succeedingly became swell,round,shrink,gathered into syncytial body and even detached from the cell bottle.IFA showed that after inoculation,the virus began to proliferate at 12 h,reached the replication peak at 36 h,then the proliferation ability decreased and tended to be steady at 60 h.Growth curves showed that SHpd/2012 proliferated at high speed within the first 24-60 h and with a decreasing trend 60 hours after inoculation.Besides,we expressed S protein,which made a foundation for studying the interaction between PEDV and its receptor and pathogenesis of the virus.