鳜鱼病毒病原及细胞感染特性的研究

Study of the Siniperca chuatsi Viral Pathogen and its Infectious Characteristics in Fish Cells

在患暴发流行性发病鳜鱼的脾、肾、肝、鳃组织细胞的超薄切片中观察到直径为 12 0 150nm的二十面体病毒粒子 ,其截面呈六角形 ,有囊膜。将病鱼内脏与 10倍体积的PBS缓冲液制成组织匀浆液 ,过滤除菌后感染对数生长期的草鱼肾细胞。结果表明 ,该感染液在 2 8℃条件下 ,培养 36 4 8h ,生长旺盛的草鱼肾细胞开始出现CPE ,5 7d感染的单层细胞逐渐脱落死亡。该细胞病毒悬液具有感染性 ,可持续稳定地进行传代培养。感染细胞的病毒培养液 ,经差异离心纯化 ,负染制片 ,电镜下观察到大量的病毒粒子 ,对照培养物中未发现任何病毒颗粒。

An icosahedral virus was found in the ultra thin section of spleen, kidney, liver, gill of the outbreak infectious Siniperca chuatsi by electron microscope. The virions of section show hexagon, about 125 150 nm in diameter, and have envelope outside. Cell culture indicated that CIK cell was sensitive to the virus, which was isolated from the internal organs of Siniperca chuatsi. The CPE can be seen after being infected for 36 48 hours at 28℃. Cell cultured virus suspension is infectious and can be subcultured steadily in CIK cell line. Many virions isolated by different centrifugation from infected cell culture can be observed by negative stained electron microscope method, but no virions were detected in the control samples.