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目的改进水痘-带状疱疹减毒活疫苗毒种(OKa株)培养方法,提高毒种的均一性及稳定性,延长保存时间。方法分别采用原培养方式(2 L方瓶培养法)和改进的培养方式(40层细胞工厂)制备水痘-带状疱疹病毒OKa株工作种子批,检测病毒滴度及-65℃以下保存1、3、6、12个月的稳定性,并用该种子批分别制备3批水痘减毒活疫苗后,进行疫苗成品的全面检定。结果 2 L方瓶培养法和40层细胞工厂制备的水痘-带状疱疹病毒工作种子批OKa-2015[1]C和OKa-2015[2]C的病毒滴度分别为6.1和6.0 lg PFU/ml;无菌试验、支原体检查、鉴别试验、外源因子检查结果均符合《中国药典》三部(2015版)相关要求;-65℃以下保存12个月,滴度均无明显下降。两种培养方式各制备的3批疫苗成品的各项检定结果均符合《中国药典》三部(2015版)相关要求,各批疫苗的病毒滴度及37℃热稳定性差别不明显。结论 40层细胞工厂制备水痘-带状疱疹病毒毒种(OKa株),-65℃以下长期保存具有稳定性,该培养方式可提高毒种的均一性,降低微生物污染风险。
Objective To improve the culture method of varicella - zoster attenuated live vaccine strain (OKa strain), to improve the uniformity and stability of the virulent species and prolong the storage time. Methods The seedlings of varicella - zoster virus OKa strain were prepared by using the original culture method (2 L square bottle culture method) and the improved culture method (40 layer cell factory) respectively. The virus titers were measured and stored below -65 ℃. 3,6,12 months of stability, and with the seed batch separately prepared three batches of live attenuated varicella vaccine, the vaccine for the full test. Results The viral titers of OKa-2015 [1] C and OKa-2015 [2] C vaccines were 6.1 and 6.0 lg PFU / kg in the 2 L bottle culture and the 40-cell factory, respectively ml. The results of sterility test, mycoplasma test, identification test and exogenous factor test all met the requirements of the third edition of Chinese Pharmacopoeia (2015 edition). No significant drop in titers was observed at -65 ℃ for 12 months. The test results of the three batches of vaccine products prepared by the two cultivation methods all met the requirements of the third part of the Chinese Pharmacopoeia (2015 edition), and the virus titer and the thermal stability at 37 ° C of the vaccines did not differ significantly. Conclusion The 40-layer cell factory was used to prepare varicella-zoster virus strain (strain OKa), with long-term storage stability below 65 ° C. This culture method can improve the uniformity of virus species and reduce the risk of microbial contamination.