肾综合征出血热疫苗后备筛选毒株在Vero细胞上的适应传代及其抗原性研究

Passage adaptability of candidate vaccine strains fro hemorrhagic fever with renal syndrome in Vero cells and their antigenicity

〔目的〕将肾综合征出血热疫苗后备毒株适应于Vero细胞 ,观察病变情况 ,并对其抗原性和繁殖特性进行研究。〔方法〕将新分离的汉坦病毒ZJ2、ZJ4、ZJ7株和汉城病毒ZJ5株在Vero细胞上进行连续传代 ,采用间接免疫荧光法(IFA)、反向被动血凝试验 (RPHA) ,研究传代后毒株的病毒滴度、抗原量以及细胞的病变情况 ,同时将传代后的病毒接种敏感动物 ,观察动物的发病情况和毒株的繁殖特性。〔结果〕经过 5次连续传代 ,四株病毒在Vero细胞上均显示出良好的适应性 ,从第二代开始病毒滴度稳定在 6.0 0LgTCID5 0 ml以上 ,第五代时最高达 7.5 0LgTCID5 0 ml ,第三代毒株抗原量均已达到 1:12 8,ZJ7株抗原量最高时达 1:768,四株毒株感染Vero细胞 ,连续观察 12d不见细胞产生病变 ,第五代细胞病毒液接种敏感动物 ,9d后动物发病甚至死亡。〔结论〕四株病毒已适应于Vero细胞 ,且具有病毒滴度高、毒力强和抗原性良好的特性 ,是Vero细胞肾综合征出血热灭活疫苗良好的后备筛选毒株。

Objective To adapt vaccine strains of hemorrhagic fever with renal syndrome (HFRS) to Vero cells and study their antigenicity and propagating features.〔Methods〕 The viral strains ZJ2,ZJ4,ZJ7 (Hantavirus,HIN) and ZJ5 (Seoul virus,SEO) were continuously propagated on Vero cells and the antigenicity of viral multiplication,viral titers and the amounts of viral antigen were studied after serial passages.〔Results〕 The strains ZJ2,ZJ4,ZJ7 and ZJ5 adapted to Vero cells and stably grew on the cells with high titers.There was no cytopathogenic effect (CPE) 12 d later.The Mongolian gerbil diseased or died 9 d after inoculation of the viral strains.〔Conclusions〕 the 4 candidate strains successfully adapt to Vero cells and have high titers and antigenicity.Therefore,they are feasible for preparation of HFRS vaccine in Vero cells.