水痘疫苗生产工艺变更前后Oka株病毒基因序列及疫苗质量分析  

Analysis of Oka strain gene sequence and varicella vaccine quality before and after production process change

作　　者：袁萍 谢蕾[2] 王月红 黄帼英 孙范玉 刘朝阳[3] 周翎乔[2] 马雷钧[3] 石玮[3] 王亮[2] 朱为 

机构地区：[1]上海生物制品研究所有限责任公司第三研究室,200051 [2]上海生物制品研究所疫苗三室,200051 [3]上海生物制品研究所质量检定室,200051

出　　处：《国际生物制品学杂志》2017年第5期209-213,共5页International Journal of Biologicals

摘　　要：目的 通过对采用细胞工厂工艺进行细胞培养前后的Oka株病毒基因序列和水痘疫苗质量进行分析比较，评价生产工艺变更对水痘疫苗质量的影响。方法 从水痘疫苗原液提取病毒基因组，采用PCR法扩增片段，通过对PCR产物测序或克隆质粒测序并拼接，获得Oka株的病毒全基因序列，并与GenBank中相关序列比对。对工艺变更前后生产的疫苗进行病毒滴度、牛血清白蛋白残留量和抗生素残留量比较。结果 工艺变更前后的疫苗病毒基因序列一致，检测到33个单核苷酸多态性位点为亲本型和疫苗型碱基共存，具有典型的Oka株特征，没有GenBank或文献报道的新突变。工艺变更前后生产疫苗的平均病毒滴度分别为5.00和5.02 lg蚀斑形成单位/ml，两者间的差异无统计学意义（t＝1.1，P＝0.26），且各项关键指标均符合企业标准和药典要求。结论 将细胞工厂引入生产工艺后，水痘疫苗具有与原工艺生产的制品相似的安全性和质量一致性。Objective To evaluate the impact of production process change on the quality of varicella vaccine through analyzing the gene sequence of varicella-zoster Oka strain and the quality of varicella vaccine before and after application of cell factory in cell culture. Methods The viral DNA were extracted from bulk of varicella vaccine and fragments were amplified by PCR. The whole gene sequence of Oka strain was obtained by DNA sequencing with PCR products or recombinant plasmids and compared to the relevant sequences in GenBank. The virus titers, residual bovine serum albumin and residual antibiotics between vaccines before and after production process change process change. Results The genetic sequence between vaccines before and after production process change was consistent, which had typical characteristics of Oka strain, and no new mutations were found. 33 single nucleotide polymorphism loci were found with a mixture of both parental and vaccine-type bases. The mean virus titers of vaccine before and after production process change were 5.00 and 5.02 lg plaque-forming unit/ml, respectively, and there was no statistical significance between them （t=1.1,P=0.26）. The key quality indexes of the vaccine before and after the process change all met the enterprise standards and pharmacopoeia requirements. Conclusion After introducing the cell factory into the production process, the varicella vaccine has similar safety and quality consistency with the product prepared by original process.

关 键 词：水痘疫苗 疱疹病毒3型 人 序列分析 Oka株 

分 类 号：R927[医药卫生—药学]