轮状病毒LLR株培养条件的优化  

Optimization of culture conditions of rotavirus strain LLR

作　　者：庞兴 马金霞 刘淦 张永鹏 魏邦孝 郭敏 李佳林 鱼轲 PANG Xing;MA Jin-xia;LIU Gan;ZHANG Yong-peng;WEI Bang-xiao;GUO Ming;LI Jia-lin;YU Ke(The First Department of Vaccine,Lanzhou Institute of Biological Products Co.,Ltd.,Center for Gansu Provincial Vaccine Engineering Research,Lanzhou 730046,Gansu Province,China)

机构地区：[1]兰州生物制品研究所有限责任公司疫苗一室甘肃省疫苗工程技术研究中心,甘肃兰州730046

出　　处：《微生物学免疫学进展》2021年第3期30-34,共5页Progress In Microbiology and Immunology

摘　　要：目的为提高轮状病毒滴度,对病毒培养过程中的关键因素如感染复数(multiplicityofinfection,MOI)、胰蛋白酶浓度及维持液pH进行优化,为口服轮状病毒活疫苗生产提供数据依据。方法将轮状病毒LLR株分别以MOI0.005、0.01、0.02、0.04和0.08接种牛肾细胞后收获病毒;以MOI0.02接种牛肾细胞,分别加入含1、2、3、4、5和7μg/mL胰蛋白酶的MEM培养液后收获病毒;以MOI0.02接种牛肾细胞,分别加入pH为6.0、6.5、7.0、7.5、8.0及8.5的MEM培养液后收获病毒。分别在24、48、72及96h取样,检测病毒滴度。结果以MOI0.02接种牛肾细胞后滴度最高,为7.5lgCCID_(50)/mL;当维持液中胰蛋白酶质量浓度为4μg/mL时滴度最高,为7.2lgCCID_(50)/mL;当pH为6.5及7.0时滴度最高,均为7.5lgCCID_(50)/mL。结论通过对牛肾细胞培养轮状病毒关键条件的优化,获得了提高病毒滴度的有效方法,为口服轮状病毒活疫苗生产过程控制提供数据依据。Objective To improve the titer of rotavirus, the key factors during the virus culture such as multiplicity of infection(MOI), trypsin concentration and pH of maintenance medium were optimized. Methods Bovine kidney cells were inoculated with rotavirus strain LLR at different MOIs of 0.005, 0.01, 0.02, 0.04 and 0.08, respectively. Bovine kidney cells were inoculated with LLR strain at a MOI of 0.02 and cultured in MEM medium containing 1, 2, 3, 4, 5 and 7 μg/mL trypsin, respectively. Bovine kidney cells were inoculated with LLR strain at a MOI of 0.02, and cultured in MEM medium at pH 6.0, 6.5, 7.0, 7.5, 8.0 and 8.5, respectively. Virus titers were detected at 24, 48, 72 and 96 hours. Results The highest virus titer(7.5 lgCCID50/mL) was obtained when bovine kidney cells were inoculated with LLR at a MOI of 0.02. The highest virus titer(7.2 lgCCID_(50)/mL) was obtained when the concentration of trypsin in maintenance medium was 4 μg/mL. The highest virus titer(7.5 lgCCID_(50)/mL) was obtained when the pH of MEM medium was 6.5 and 7.0. Conclusion An effective method to improve the virus titer was obtained by optimizing the cultural conditions of LLR strain in bovine kidney cells, which provides a basis for the quality control of the production process of oral rotavirus vaccine.

关 键 词：牛肾细胞 轮状病毒LLR株 感染复数 胰蛋白酶浓度 PH 病毒滴度 

分 类 号：R373.25[医药卫生—病原生物学]