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将不同培养时间及不同传代次数的病毒用PBS稀释后,感染健康草鱼(5～15 cm),取感染后具有典型症状的草鱼肾脏组织液,对健康草鱼进行重复感染,对其致病和致死率进行统计,并用抗草鱼呼肠孤病毒的单克隆抗体5F11进行间接免疫荧光检测.结果显示长期培养(>8 d)的病毒外衣壳脱落,其对草鱼的致病及致死率与完整衣壳感染无显著差异;传代不同次数(1～50代)的病毒对草鱼的致病及致死率无显著差异;复染的草鱼其致病及致死率与初染无显著性差异,间接免疫荧光检测复染的草鱼肾脏冷冻切片能观察到明亮的点状黄绿色荧光.即体外长期培养能导致外衣壳脱落,但这对草鱼呼肠孤病毒的毒力无影响,且体外传代(1～50代)也不影响草鱼呼肠孤病毒的毒力.
After being infected with healthy grass carp (5-15 cm), the virus with different culture times and different passages was diluted with PBS, and the grass carp kidney tissue fluid with typical symptoms after infection was infected. The pathogenicity and lethality of the grass carp The results of indirect immunofluorescence assay showed that the outer shell of the virus was shed in long-term culture (> 8 d), and the pathogenicity and lethality of grass carp was the same as that of the complete capsid infection There was no significant difference in the pathogenicity and lethality of the grass carp from different passages (from 1 to 50 passages). There was no significant difference in the pathogenicity and lethality between the infected and inoculated grass carp. The indirect immunofluorescence assay Frozen grass carp kidney sections can be observed bright spot-like yellow-green fluorescence, that is, long-term in vitro culture can lead to shed shell, but this grass carp reovirus virulence had no effect, and in vitro passage (1 to 50 generations) Does not affect the toxicity of grass carp reovirus.