流感病毒在Vero细胞上的适应性

目的研究WHO指定流感疫苗生产用毒株在Vero细胞上的适应性,为用Vero细胞制备流感疫苗奠定基础。方法优化不同培养基、胰酶浓度、pH值等培养病毒的条件及收毒时间,将流感疫苗生产用毒株在Vero细胞上连续传代,并进行血凝效价检测及RT-PCR分析。结果在病毒维持液为F12+DMEM、pH为7.5、胰酶含量为1.5μg/ml、培养3 d收获病毒时,可获得较高的病毒血凝效价,连续传4代,病毒血凝效价降为0,RT-PCR检测结果为阴性。结论WHO推荐的流感疫苗生产用毒株在Vero细胞上连续传代,病毒血凝效价逐渐降低。

Adaptability of Influenza Virus in Vero Cells

Objective To study the adaptability of influenza virus strain recommended by WHO for production of vaccine in Vero cells and lay a foundation of preparation of influenza vaccine with Vero cells.Methods The condition for culture of influenza virus,such as medium,trypsin concentration,pH value and time for virus harvest,were optimized.The influenza virus strain for production of vaccine was subcultured continuously in Vero cells for determination of HI titer and for PT-PCR analysis.Results The optimal medium,trypsin concentration,pH value and time for virus harvest for culture of influenza virus were F12+DMEM,1.5 μg/ml,7.5 and 3 d respectively.The virus cultured under the optimized condition reached a high HI titer.However,after subculture in Vero cells for 4 passages,the HI titer of virus decreased to 0,and the result of RT-PCR was negative.Conclusion The HI titer of influenza virus strain recommended by WHO for vaccine production decreased gradually after subculture in Vero cells.