水泡性口炎病毒在草鱼肾细胞中的复制和病变效应技术报告.docVIP

 水泡性口炎病毒在草鱼肾细胞中的复制和病变效应 摘要：目的 观察水泡性口炎病毒（Vesicular stomatitis virus ,VSV）感染草鱼肾细胞（CIK细胞）所引起细胞病变效应并检测其在细胞中的复制。方法 用VSV病毒印第安纳（VSV Indiana）CIK细胞，分别在12h、24h、36hRNA，通过RT-PCR方法检测病毒在细胞中的复制。结果 受VSV病毒感染后的CIK细胞圆缩、聚集、脱落，形成包涵体，随着时间的延长该现象逐渐加剧。RT-PCR结果显示出现了预期的200 bp左右的条带。结论 VSV病毒引起了CIK细胞的病变，并在CIK细胞中进行了复制。 关键词：VSV，CIKVesicular Stomatitis Virus in Ctenopharyngodon Idellus Kidney Cells Replication and Cytopathic Effect GAO Xia-Ting (Qiuzhen School of Huzhou Teachers College, Huzhou 313000) Abstract: Objective We observed cytopathic effect in Ctenopharyngodon idellus kidney cells (CIK cells) induced by Vesicular stomatitis virus (VSV) and detected VSV’s replication in it. Methods CIK cells were infected by VSV Indiana strains. After 12 h, 24 h, 36 h, the cell cytopathic effect was observed. When a wide range of cytopathic effect appeared, the virus’s RNA would be extracted and detected by RT-PCR. Results CIK cells with VSV became round shrinkage, got together, shaded, and formed inclusion body. With the extension of time，it increased. RT-PCR result showed that the expected bands of 200bp were amplified with templates of virus’s RNA. Conclusion Cytopathic effect appeared in CIK cells with VSV. And VSV could reproduce in CIK cells. Key words: VSV, CIKcells, cytopathic effect, replication 目 录 1 引言………………………………………………………………………………………1 2 实验材料和方法…………………………………………………………………………2 2.1材料……………………………………………………………………………2 2.1.1 实验细胞 2.1.2 主要化学药品和试剂2.1.3 主要仪器与设备 2.1.4 试剂配制2.2 方法与步骤…………………………………………………………………………4 2.2.草鱼的肾细胞(CIK细胞)培养 2.2.2 VSV病毒的复壮与效价检测 2.2.3 细胞病变效应 2.2.4 病毒RNA提取及RT-PCR测定草鱼肾细胞(CIK细胞)中的3 结果与分析 3.1 正常草鱼的肾细胞（CIK细胞）培养特征…………………………………………11 3.3 VSV病毒感染草鱼肾细胞（CIK细胞）不同时间后产生的细胞病变效应……11 3.4 RT-PCR检测VSV病毒在CIK细胞中的复制…………………………………12 3.5空斑试验……………………………………………………………………………13 4 讨论………………………………………………………………………………………1 4.1 VSV病毒对不同细胞系引起病变效应……………………………………………13 4.2 VSV病毒对Vero细胞、3T3细胞、CIK细胞的空斑实验结果的比较…………14 4.3 VSV病毒的检测方法………………………………………………………………15