草鱼呼肠孤病毒生长特性研究

Research on Growth Characteristics of Grass Carp Reovirus

以草鱼呼肠孤病毒(GCRV)873毒株为试验材料,建立了GCRV荧光定量PCR检测方法;用GCRV873株感染CIK细胞及草鱼,通过对病毒RNA复制水平、子代病毒含量以及细胞病变等情况进行监测,研究其在CIK细胞及草鱼体内的生长特性。在CIK细胞中,GCRV感染后12 h即可检测到子代病毒,36 h时,所有细胞被感染;72 h时,病毒滴度达到最高,为10^(6.75) TCID_(50)/mL。在草鱼的肝、脾、肾中,均能检测到GCRV RNA,其中肾脏中含量最高,其次是肝脏,脾脏中最少。本试验为研究GCRV致病机理及制备细胞灭活疫苗奠定了试验基础。

Taking Grass carp reovirus（GCRV）873 strain as the test material,a GCRV fluorescence quantitative PCR detection method was established. First,CIK cells and grass carps were infected by 873 strain of GCRV,then the replication level of virus RNA,subgeneration virus content and cytopathic conditions were monitored,and the growth characteristics of GCRV in CIK cells and grass carps were studied. In CIK cells,the subgeneration virus could be detected at 12 h after GCRV infection. After 36 h,almost all cells were infected by viruses. After 72 h,the virus titer reached the top,which was nearly 106.75 TCID50 /mL. Besides,GCRV RNA could be detected in both liver,spleen and kidney of grass carps. Among them,GCRV RNA in kidney was the most,followed by in liver,and in spleen it was the least. The experiment laid foundation for studying the pathogenesis of the virus and preparing the inactivated vaccines.