不同感染方式对狂犬病病毒在人二倍体细胞中增殖的影响  

The effects of different infection methods on rabies virus proliferation in human diploid cells

作　　者：甄祖刚 周荔葆 王野 刘春雨 张怡轩[1] ZHEN Zugang;ZHOU Libao;WANG Ye;LIU Chunyu;ZHANG Yixuan(School of Life Science and Biopharmaceutics,Shenyang Pharmaceutical University;Liaoning Chengda Biological Co.,Ltd.)

机构地区：[1]沈阳药科大学生命科学与生物制药学院,辽宁沈阳110016 [2]辽宁成大生物股份有限公司,辽宁沈阳110179

出　　处：《沈阳药科大学学报》2024年第2期177-181,共5页Journal of Shenyang Pharmaceutical University

基　　金：辽宁省“兴辽英才计划”项目(XLYC1902072)。

摘　　要：目的考察带毒传代感染、直接吸附感染、混种传代感染三种方式感染人二倍体细胞对狂犬病病毒增殖的影响。方法不同方式感染后,经过细胞工厂生产,获取狂犬病病毒收获液,经过滤澄清、浓缩、纯化、灭活制备疫苗原液。根据《中华人民共和国药典》(2020版)检测方法检测收获液病毒滴度及原液抗原含量。结果带毒传代组的收获液病毒滴度为(6.40±0.33)lgLD50·mL^(-1),原液抗原含量为(34.8±1.5)IU·mL^(-1);混种传代感染组的收获液病毒滴度为(6.35±0.18)lgLD50·mL^(-1),原液抗原含量为(34.3±2.1)IU·mL^(-1);直接吸附感染组收获液病毒滴度为(4.71±0.39)lgLD50·mL^(-1),原液抗原含量为(12.6±2.0)IU·mL^(-1)。结论试验证明狂犬病病毒PV株采用混种传代感染的方式感染人二倍体细胞MRC-5细胞,病毒增殖效果良好,毒种代次明确,工艺操作简单,更适合于狂犬疫苗的生产。Objective To investigate the effect of infection of human diploid cells by virus-carrying passage,direct adsorption and mixed passage on the proliferation of rabies virus.Methods After infection with different methods,the rabies virus harvesting solution was obtained from the manufacturing in the cell plant,and the vaccine stock solution was prepared by filtration and clarification,concentration,purification and inactivation.The virus titer of the harvesting solution and antigen content of the stock solution were determined using the assay method of Chinese Pharmacopoeia(2020).Results The virus titers of the harvesting solutions for virus-carrying passage infection group,mixed passage infection group and direct adsorption infection group were(6.40±0.33),(6.35±0.18)and(4.71±0.39)lgLD_(50)·mL^(-1),respectively.The antigen contents of the stock solutions were(34.8±1.5),(34.3±2.1)and(12.6±2.0)IU·mL^(-1),respectively.Conclusion After the rabies virus PV strain infected human diploid cells MRC-5 cells by mixed passage,good virus proliferation was shown with a clear viral strain generation and a simple process.Therefore,this method was more suitable for the manufacturing of rabies vaccine.

关 键 词：狂犬病病毒 人二倍体细胞 感染方式 抗原含量 

分 类 号：R181[医药卫生—流行病学]