麻疹、腮腺炎、风疹和水痘联合减毒活疫苗的研制

目的 建立麻疹、腮腺炎、风疹和水痘(measles,mumps,rubella and varicella,MMRV)联合减毒活疫苗的生产工艺和检定方法.方法 采用麻疹病毒沪-191株、腮腺炎病毒S79株、风疹病毒BRDⅡ株、水痘-带状疱疹病毒北京84-7株,在原代鸡胚细胞或人胚肺二倍体细胞2BS株中制备高滴度疫苗病毒原液.观察4种原液按不同比例稀释配制后病毒的滴度变化和相互干扰现象,确定MMRV疫苗中4种原液的配制比例,并筛选适宜保护剂,建立最佳冻干工艺.同时,建立MMRV联合减毒活疫苗的检定方法.采用t检验对结果进行比较.结果 选择最佳配制比例、保护剂和冻干工艺制备出连续多批MMRV疫苗,按国家药典要求检定全部合格.其中连续3批疫苗经国家检定机构检定合格:麻疹病毒基础滴度和37℃放置7d后的滴度分别≥3.9和≥3.5 lg半数细胞培养感染量(50％ cell culture infective dose,CCID50)/ml,腮腺炎病毒≥5.0和≥4.7 lgCCID50/ml,风疹病毒≥5.0和≥4.8 lgCCID50/ml,水痘病毒≥4.5和≥4.4 lg噬斑形成单位/ml.用建立的方法检测MMRV疫苗,结果4种病毒滴度实测值与理论值之间的差异无统计学意义(t值为0.149～1.838,P值均＞0.05).结论 建立了稳定、可行的MMRV疫苗生产工艺和检定方法.

Preparation of a live attenuated measles,mumps, rubella and varicella combined vaccine

Objective To establish the production process and titration method of a live measles,mumps,rubella and varicella (MMRV) combined vaccine.Methods Four high-titer virus bulks were prepared using measles virus strain Shanghai-191,mumps virus strain S79,rubella virus strain BRD Ⅱ and varicella-zoster virus strain Beijing 84-7 cultured in primary chicken embryo cell or human embryo lung diploid cell 2BS.The titer change and mutual interference of 4 kinds of viruses were observed after preparation with different dilution ratios.The appropriate protective agent was screened and the best freeze-drying condition determined.Meanwhile,the titration method of live MMRV vaccine was established.The results were compared with t-test.Results Several consecutive batches of MMRV vaccine were prepared with the best dilution ratio,protective agent and freeze-drying process and met the requirements in Chinese pharmacopoeia.Among which,3 consecutive batches were determined by national verification institution.The virus titers before and after 7 days at 37 ℃ were as follows:measles ≥3.9 and ≥3.5 lg 50％ cell culture infective dose (CCID50)/ml,mumps ≥5.0 and ≥4.7 lgCCID50/ml,rubella ≥5.0 and ≥4.8 lgCCID50/ml,varicella ≥4.5 and ≥4.4 lg plaque-forming unit/ml.Determined by the established method (t =0.149-1.838,all P ＞0.05),there was no significant difference between the measured and theoretical titers of 4 kinds viruses in MMRV vaccine.Conclusion The production process and titration method established are stable and feasible.