为研究新型鸭呼肠孤病毒(novel duck reovirus,NDRV)在BHK21细胞系中的增殖规律,将NDRV野毒株TH11接种BHK21细胞系,连续传代3次后,观察病毒对细胞的致病变效应(CPE),并对病毒核酸及其蛋白的表达和病毒增殖特性进行了鉴定。结果显示,NDRV TH11株能够在BHK21细胞中有效增殖,并产生以细胞巨融合为主的典型CPE,病毒蛋白在细胞中也获得了良好表达,并与σC蛋白特异性抗体发生反应。一步生长曲线显示,TH11毒株感染BHK21细胞后第0~12小时为潜伏期,第12小时开始增殖,第12~36小时进入快速增殖期,第48小时病毒效价即达到最高,即TCID50为106.57/0.1mL,随后病毒的增殖随细胞的崩解而降低。上述结果为进一步研究NDRV的致病机理和研制细胞灭活疫苗奠定了基础。
To explore the proliferation characteristics of novel duck reovirus(NDRV)in BHK21 cells,BHK21 cells were inoculated with NDRV TH11 strain.After passaged three times,CPE could be observed clearly at hour 12 post-infection.The results of RT-PCR,IFA and Western-blot showed that viral NDRV-σC was expressed well in BHK21 cells,and reacted to the specific antibody against NDRV-σC.One step growth curve revealed that NDRV TH11 strain began to proliferate at hour 12,and reached a rapid proliferation stage during hour 12 to 36post-infection.The virus titer of NDRV TH11 strain reached peak at hour 48.In summary,the above-mentioned results provide some valuable information for further exploring the pathogenesis mechanism of NDRV and vaccine development.