兔出血症病毒体外复制的研究  

Studies on Replication of Rabbit Hemorrhagic Disease Virus (RHDV) in Vitro

作　　者：罗经[1] 杨学楼 刘虹[1] 严银钫[1] 王汉中[1] 孙松柏[1] 

机构地区：[1]中国科学院武汉病毒研究所,武汉430071

出　　处：《中国病毒学》1992年第3期343-350,共8页Virologica Sinica

基　　金：国家自然科学基金

摘　　要：在几种动物细胞上,采用同步感染的方法研究了兔出血症病毒(RHDV)的复制特性。病毒感染细胞后(PI)48—72小时可观察到明显的细胞病变,血凝效价可增高5—10倍。病毒对细胞传代代数不同,敏感性也不同。在兔婴肾(RK)和兔婴肺(RL)细胞上以4—8代最为敏感。采用免疫荧光染色法,经病毒感染48—72小时的细胞中可观察到特异性荧光。细胞增殖的病毒经PEG-DS浓缩,Sepharose 4B柱层析提纯后,在电镜下可观察到完整的病毒粒子,将此病毒回接健康实验兔可致100％死亡。免疫双扩散和免疫电泳试验表明,细胞增殖的病毒抗原与来自病兔肝的RHDV抗血清之间产生明显的沉淀带。SDS-PAGE分析病毒获得四条多肽,其分子量大小与病兔肝组织提取的病毒蛋白多肽分子量相比略有差异。In this paper the characteristics of replication of RHDV in the RL and RK cells have been studied by using synchronous infective method. The cytopathology effect can be observed at 48—96 hrs after viruses infection. HA titer of viral increased 5—10 times as high as those HA titers of viruses infection at 72 hr. The sensitivity of RHDV to passage cells are different, it is most sensitive for 4—8 generation. Special fluorescence in the ceils can be seen at 18—72 hrs using immunofluorescence stain. Viruses of multiplication in cells were precipitated with PEG-DS, and purified with Sepharose 4B chromatography. Intact virions can be observed under electronmicroscope. Healthy rabbits were infectious with pure virions, resulting in death of rabbits by 100％. Clear immuno-sedimentation band can be produced between antigens obtained from cell cultures with antisera of RHDV using ID and CIE tests. The pure virions of cell cultures resolved into four polypeptides and with molecular weight of 65kd, 52kd, 43kd and 36kd respectively was seen after coomassie blue staining following SDS-PAGE. The molecular weight of Vp2, Vp3, Vp4 polypeptide showed identical with Vp2, Vp3, Vp4 polypeptide of virions of rabbit liver tissue but molecular weight of Vp1 polypeptide were slight different each other.

关 键 词：兔出血症病毒 血凝效价 复制 

分 类 号：S852.65[农业科学—基础兽医学]