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猪流行性腹泻病毒SHpd/2012株侵染Vero细胞特性分析
引用本文:谢洋洋,丁思嘉,郑煜明,罗依然,杨志彪.猪流行性腹泻病毒SHpd/2012株侵染Vero细胞特性分析[J].上海交通大学学报(农业科学版),2018(2):16-21.
作者姓名:谢洋洋  丁思嘉  郑煜明  罗依然  杨志彪
作者单位:上海交通大学农业与生物学院上海市兽医生物技术重点实验室
基金项目:国家自然科学基金(31472211);国家重点研发计划(2016YFD0500100)
摘    要: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PEDVSHpd/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12h病毒开始增殖,在接毒后36h时感染细胞数量最多,其后病毒增殖速度逐步下降,60h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24~60h的毒价一直处于较高水平,60h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。

关 键 词:PEDV(猪流行性腹泻病毒)    侵染细胞    S基因    真核表达
收稿时间:2017/5/30 0:00:00

Characteristics of vero cells infected by SHpd/2012 strain of porcine epidemic diarrhea virus
XIE Yang-yang,DING Si-ji,ZHENG Yu-ming,LUO Yi-ran and YANG Zhi-biao.Characteristics of vero cells infected by SHpd/2012 strain of porcine epidemic diarrhea virus[J].Journal of Shanghai Jiaotong University (Agricultural Science),2018(2):16-21.
Authors:XIE Yang-yang  DING Si-ji  ZHENG Yu-ming  LUO Yi-ran and YANG Zhi-biao
Institution:Shanghai Key Laboratory of Veterinary Biotechnology,School of Agriculture and Biology, Shanghai Jiaotong University,Shanghai 200240,China,Shanghai Key Laboratory of Veterinary Biotechnology,School of Agriculture and Biology, Shanghai Jiaotong University,Shanghai 200240,China,Shanghai Key Laboratory of Veterinary Biotechnology,School of Agriculture and Biology, Shanghai Jiaotong University,Shanghai 200240,China,Shanghai Key Laboratory of Veterinary Biotechnology,School of Agriculture and Biology, Shanghai Jiaotong University,Shanghai 200240,China and Shanghai Key Laboratory of Veterinary Biotechnology,School of Agriculture and Biology, Shanghai Jiaotong University,Shanghai 200240,China
Abstract: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.
Keywords:PEDV(porcine epidemic diarrhea virus)  infect cells  S gene  expression of protein
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