水痘-带状疱疹病毒(VZV84-7株)在人二倍体细胞(2BS株)中感染复数的分析  

Analysis of multiplication of infection of varicella-zoster virus 84-7 strain in 2BS strain of human diploid cells

作　　者：侯良玉[1,2] 麻广[2] 丁秀红[2] 黄林[2] 牟鑫尧 李立[2] 刘亚宇[2] 李永忠[2] 刘晔[2] 

机构地区：[1]四川大学华西医院生物治疗国家重点实验室,四川成都610041 [2]成都生物制品研究所有限责任公司,四川成都610023

出　　处：《中国生物制品学杂志》2014年第7期878-880,共3页Chinese Journal of Biologicals

摘　　要：目的探讨水痘-带状疱疹病毒（VZV84-7株）在人二倍体细胞（2BS株）上的感染活性,确定感染复数（MOI）。方法使用100 ml小方瓶培养人二倍体细胞（2BS株）,待长成致密单层后,接种病毒滴度为5.5 lgPFU/ml的水痘-带状疱疹病毒（VZV84-7株）,每瓶接种量分别为0.02、0.1、0.3、0.5、0.8、1、1.5和2 ml,于显微镜下连续观察细胞病变情况及病变比例,并连续记录收获时间;采用蚀斑法测定病毒滴度。结果当病毒接种量为0.02 ml时,细胞大部分生长正常,呈极性排列,无形态典型、广泛的病变,通过延长收获时间也无法获得形态典型、广泛的病变;当病毒接种量不低于0.1 ml时,呈典型、广泛的病变,随着病毒接种量的增加,收获时间逐渐缩短;仅病毒接种量为0.02 ml组细胞的病毒滴度低于3.7 lgPFU/ml;确定MOI在0.004 0~0.019 9之间,均可制备出质量稳定、符合成都生物制品研究所有限责任公司《水痘减毒活疫苗制造及检定规程》要求的毒种。结论确定了水痘-带状疱疹病毒（VZV84-7株）感染人二倍体细胞（2BS株）合适的MOI,为水痘疫苗生产工艺的优化提供了参考依据。Objective To investigate the infectious activity of varicella-zoster virus（VZV）84-7 strain in human diploid cells（2BS strain）and determine the multiplication of infection（MOI）. Methods 2BS cells were cultured in 100 ml small square vase,and inoculated with VZV84-7 strain at a titer of 5. 5 lgPFU / ml after formation of a dense monolayer. The inoculum sizes in various vases were 0. 02,0. 1,0. 3,0. 5,0. 8,1. 0,1. 5 and 2. 0 ml respectively. CPEs and their percentages were observed under microscope,and the harvest time was recorded continuously. The virus titer was determined by plaque assay. Results When the inoculum size was 0. 02 ml,most of 2BS cells grew normally and in a polar arrangement. No typical and extensive CPEs were observed,even after the harvest time was delayed. However,when the inoculum size was not less than 0. 1 ml,typical and extensive CPEs were observed,and the harvest time decreased gradually with the increasing inoculum size. Only the titer of virus at an inoculum size of 0. 02 ml was less than 3. 7 lgPFU / ml.The optimal MOI was determined as 0. 004 0 ~ 0. 019 9,at which the prepared virus seeds showed stable quality and met the Requirements for Live Attenuated Varicella Vaccine issued by Chengdu Institute of Biological Products Go.,Ltd.as a standard for license. Conclusion The optimal MOI for infection of 2BS cells with VZV84-7 strain was determined,which provided a reference for optimization of production procedure for varicella vaccine.

关 键 词：水痘-带状疱疹病毒 二倍体细胞系 感染复数 

分 类 号：R373.11[医药卫生—病原生物学] R392.33[医药卫生—基础医学]