乙型脑炎减毒活疫苗生产用地鼠易感病毒ELISA检测方法的优化及验证

Optimization and validation of enzyme linked immunosorbent assay for hamster-susceptible viruses used for production of live attenuated Japanese encephalitis vaccine

目的对乙型脑炎减毒活疫苗生产用地鼠易感病毒ELISA检测方法进行优化,并对该方法进行验证。方法将ELISA方法进行优化后,检测地鼠血清中的呼肠孤病毒3型(reovirus 3,Reo3)、小鼠肺炎病毒(pneumonia virus of mice,PVM)、仙台病毒(Sendai virus,SV)和淋巴细胞脉络丛脑膜炎病毒(lymphocytic choriomeningitis virus,LCMV),验证该方法的重复性、中间精密性及特异性,并根据WHO/VSQ/97.02的要求,制定验证参数的可接受标准。结果优化后的ELISA方法具有良好的重复性、中间精密性及特异性,各项验证参数均达到预先设定的可接受标准。结论该方法可用于乙型脑炎减毒活疫苗的质量控制。

Objective To optimize and validate the enzyme linked immunosorbent assay（ELISA）for hamster-susceptible viruses used for production of live attenuated Japanese encephalitis（JE） vaccine.Methods The ELISA was optimized,and used for determination of the reovirus 3（Reo3）,pneumonia virus of mice（PVM）,Sendai virus（SV）and lymphocytic choriomeningitis virus（LCMV） in sera of hamster to validate the reproducibility,intermediate precison and specificity,based on which the acceptable criteria for validation parameters was established according to the requirements in WHO /VSQ / 97.02.Results The optimized ELISA showed high reproducibility,intermediate precison and specificity,of which the various validation parameters met the designed acceptable criteria.Conclusion The optimized ELISA may be used for the quality control of live attenuated JE vaccine.