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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 passage stability of Sabin poliovirus on Vero cells. Methods Vero cells were inoculated with Sabin strains poliovirus types Ⅰ, Ⅱ and Ⅲ at 0.01, 0.1 and 1 MOI respectively for subculture for 13 passages. The titer of the virus was determined by the method of micro cytopathic assay. The content of D antigen was determined by sandwich enzyme-linked immunosorbent assay (ELISA). The Wister rats were immunized and the immunogenicity of the virus was determined by micro-neutralization assay. Results The serotypes of type Ⅰ, type Ⅱ and type Ⅲ of Sabin strain were continuously passaged on Vero cells for 13 generations and the virus titer was basically stable. The content of D antigen showed a decreasing trend and the immunogenicity decreased rapidly. Conclusion Preparation of inactivated vaccine with Sabin poliovirus, the lower the generation pass on Vero cells, the better.