水痘-带状疱疹病毒疫苗株在Vero细胞中的传代适应

Adaptation of Varicella-Zoster Virus Vaccine Strain in Vero Cells

目的探索水痘-带状疱疹病毒(VZV)疫苗株在Vero细胞中的传代适应及大量制备抗原的可行性。方法以VZV疫苗株在Vero细胞中传代适应,并检测适应株的生物学性状,采用15L旋转培养瓶培养,大量制备VZV ELISA抗原,并检测抗原的特异性。结果VZV疫苗株在Vero细胞中传代至第9代时,CPE明显增多,病毒滴度从2～3logPFU/ml提高到不低于4·2logPFU/ml。所制备毒种的适宜MOI为0·01,在72h左右收获的病毒滴度较高。毒种在-70℃保存1年后,病毒滴度未显著降低。抗VZV阳性血清检测证实制备的抗原具有较好特异性,抗原最适稀释浓度为1:4000。结论VZV疫苗株在Vero细胞中能传代适应,并可用15L瓶旋转培养大量制备VZV抗原。

Objective To explore the feasibility of adapting varicella-zoster virus （VZV） vaccine strain in Vero cells and preparing VZV antigen in a large quantity. Methods Adapt VZV vaccine strains in Vero cells and test for the biological characters of adapted strain. Prepare VZV antigen in a large quantity by culture in 15 L spinner and test for specificity by ELISA. Results After VZV vaccine strain was subeultured in Vero cells for 9 passages, obvious CPE was observed, and the virus titer increased from 2-3 logPFU/ml to not less than 4. 2 logPFU/ml. The optimal MOI of adapted strain was 0. 01 ,and the optimal harvest time was 72 h. The virus titer of the adapted strain showed no significant decrease after storage at -70℃ for 12 months. The ELISA with VZV-positive serum proved good specificity of the prepared VZV antigen. The optimal dilution of the antigen was 1 ： 4 000. Conclusion VZV vaccine strain was adapted in Vero cells,and VZV antigen was prepared in a large quantity by culture in 15 L spinner.