新型水痘-带状疱疹病毒Oka-7S株的分离纯化及检定  

Isolation,purification and identification of a novel varicella-zoster virus Oka-7S strain

作　　者：孙跃秋 刘洪涛 姜雪鸥 李萌 魏波 陶金玉 王梦涵 周宏达 张夫坤 SUN Yueqiu;LIU Hongtao;JIANG Xueou;LI Meng;WEI Bo;TAO Jinyu;WANG Menghan;ZHOU Hongda;ZHANG Fukun(Research Department of Changchun Keygen Biological Products Co.Ltd.,Changchun 130062,Jilin Province,China)

机构地区：[1]长春祈健生物制品有限公司研究室,吉林长春130062

出　　处：《中国生物制品学杂志》2025年第2期137-142,共6页Chinese Journal of Biologicals

基　　金：吉林省科技发展计划(20200404191YY)。

摘　　要：目的分离纯化经基因改造的水痘-带状疱疹病毒(varicella-zoster virus,VZV)Oka-7S毒株,并进行检定,为新型水痘减毒活疫苗Oka-7S株毒种库的建立奠定基础。方法将Oka-7S毒株在人二倍体细胞2BS株上进行2次空斑纯化,挑选多株空斑进行扩增传代,根据细胞病变效应(cytopathic effect,CPE)及病毒滴度选择CPE典型的毒株。对分离的毒种进行病毒滴度、基因序列及蛋白检测,并按照《中国药典》三部(2020版)方法对毒株进行质量检测。结果成功筛选获得2株Oka-7S候选毒株,鉴别试验表明候选株为VZV,连续传代后病毒滴度维持在6.0 lgPFU/mL以上;Western blot检测病毒主要抗原gE蛋白正常表达,未检测到ORF7蛋白,Oka-7S毒株在传代过程中主要基因序列ORF62和编码gE的ORF68核酸序列均未发生突变;2株Oka-7S毒株无菌、支原体、外源病毒因子检测结果均为阴性。结论成功筛选出2株新型VZV Oka-7S毒株,可作为新型水痘疫苗的候选疫苗株。Objective To isolate,purify and identify the genetically modified varicella-zoster virus(VZV)Oka-7S strain,so as to lay a foundation for the establishment of a seed bank for the novel live attenuated varicella vaccine Oka-7S strain.Methods The Oka-7S strain was purified twice on 2BS human diploid cells using plaque purification method,and multiple plaque strains were selected for amplification and passage.Typical CPE strains were selected according to the cytopathic effect(CPE)and virus titer.The virus titer,gene sequence and protein of the isolated virus strains were detected,and the quality testing was performed in accordance with the methods of Chinese Pharmacopoeia(VolⅢ,2020 edition).Results Two candidate strains of Oka-7S were successfully screened,and the identification test showed that the candidate strains were VZV,of which the virus titers remained above 6.0 lgPFU/mL after serial passages.Western blot revealed that the main antigen gE protein was expressed normally,while no ORF7 protein was detected.Meanwhile,during the passage of Oka-7S strain,the main gene sequences ORF62 and ORF68 encoding gE did not mutate.The results of bacterial testing,mycoplasma testing,and exogenous viral factor testing were all negative.Conclusion Two new VZV Oka-7S strains have been successfully screened,which can be used as the candidate vaccine strains for novel varicella vaccines.

关 键 词：水痘-带状疱疹病毒 开放阅读框7 空斑纯化 

分 类 号：R373.9[医药卫生—病原生物学]