用鸡胚培养对虾白斑综合症病毒(WSSV)的研究

Propagation of white spot syndrome virus(WSSV) in embryonated eggs

为探索对虾白斑综合症病毒(WSSV)能否在鸡胚中培养,基于稳定培养体系,将WSSV接种到SPF鸡胚中.收获接种3 d后死亡的鸡胚,从中提取病毒;将其再次接种到健康鸡胚中,传代5次后,由卵黄囊膜提取病毒,将其经肌肉注射至克氏原螯虾,结果实验组克氏原螯虾100%死亡,而对照组无死亡情况.对各代鸡胚及克氏原螯虾进行PCR和电镜检测,实验组呈WSSV阳性;电镜下,死亡鸡胚细胞中可见大量正在组装的病毒粒子;表明WSSV可用鸡胚进行培养.

In order to establish a stable system of cultivating white spot syndrome virus （WSSV）, the specific pathogen free chicken embryonated eggs are inoculated with WSSV suspensions. Both the yolk sac membrane and yolk sac fluid are harvested to prepare passage inoculation suspensions. Passages 2-5 are carried out by inoculating SPF CEE via the yolk sac route. Yolk sac membrane and yolk sac fluid are detected via transmission electron microscope （TEM） observation and PCR. A great deal of WSSV particles are identified in the ultra-thin sections of experimental groups. Positive PCR results in experimental groups inoculated with WSSV and negative results in control groups are obtained. In artificial infection, WSSV isolated from the fifth passage of experimental chicken embryo results in 100% mortality to Procambarus clarkia, which confirms the success of replication of WSSV in CEE.