Sabin株脊髓灰质炎病毒在Vero细胞上的传代稳定性

Stability of Poliomyelitis Virus Sabin Strain during Subculture in Vero Cells

目的探讨Sabin株脊髓灰质炎病毒在Vero细胞上的传代稳定性。方法用Sabin株脊髓灰质炎病毒Ⅰ、Ⅱ、Ⅲ型分别以0.01、0.1和1MOI接种Vero细胞,进行传代培养,共传13代。采用微量细胞病变法测定病毒滴度;双抗体夹心法测定D抗原含量;免疫Wister大鼠,采用微量中和试验测定病毒的免疫原性。结果Sabin株脊髓灰质炎病毒Ⅰ、Ⅱ、Ⅲ型在Vero细胞上连续传13代,病毒滴度基本稳定;D抗原含量呈下降趋势;免疫原性下降迅速。结论用Sabin株脊髓灰质炎病毒制备灭活疫苗,在Vero细胞上传代的代次越低越好。

Objective To investigate the stability of poliomyelitis virus Sabin strain during subculture in Vero cells. Methods Poliomyelitis virus Sabin strain types Ⅰ, Ⅱ and Ⅲ were inoculated onto Vero cells at MOIs of 0. 01, 0. 1 and 1 separately and subcultured for 13 passages. The virus titer was determined by micro-CPE and the D antigen content by double antibody sandwich ELISA. Wister rats were immunized with the subcuhured virus, and the immunogenicity was determined by microneutralization test. Results After subculture in Vero cells for 13 passages, the titers of Sabin strain types Ⅰ , Ⅱ and Ⅲwere basically stable, while D antigen content showed a decreasing tendency, and the immunogenicity decreased rapidly. Conclusion The poliomyelitis virus Sabin strain used for production of inactivated vaccine shall be at a low passage as far as possible in Vero cells.