| 猪流行性腹泻病毒变异株CH/GX/750A/2015在Vero细胞上转瓶培养条件的优化试验 |
| |
| 引用本文： | 卢冰霞,陶明华,赵硕,秦毅斌,刘磊,陈忠伟,何颖,段群棚,梁家幸,周英宁,李斌,蒋冬福,杨思仪,赵武.猪流行性腹泻病毒变异株CH/GX/750A/2015在Vero细胞上转瓶培养条件的优化试验[J].中国畜牧兽医,2020,47(5):1506-1513. |
| |
| 作者姓名： | 卢冰霞 陶明华 赵硕 秦毅斌 刘磊 陈忠伟 何颖 段群棚 梁家幸 周英宁 李斌 蒋冬福 杨思仪 赵武 |
| |
| 作者单位： | 1. 广西兽医研究所, 广西兽医生物技术重点实验室, 南宁 530001;2. 江西省上饶市畜牧兽医科学研究所, 上饶 334000;3. 广西大学, 南宁 530005;4. 广西农业职业技术学院, 南宁 530007 |
| |
| 基金项目： | 广西科技重大专项（桂科AA17204057、桂科AA18118051）；广西自然科学基金项目（2017GXNSFBA198092）；广西兽医生物技术重点实验室开发基金课题（17-259-36-B-2、16-380-45-B-3）；广西基本科研业务费专项（桂科专项17-2） |
| |
| 摘 要： | 本研究旨在确定猪流行性腹泻病毒（PEDV）变异株CH/GX/750A/2015株在Vero细胞上转瓶培养的最佳条件，为大规模生产高效PEDV变异株疫苗提供技术支撑。试验以10 L转瓶培养病毒，对接毒时细胞维持液中胰酶浓度（0、2、4、6、8和10 μg/mL）、细胞密度（40%、60%、80%、100%、200%）、接毒剂量（MOI分别为1、0.1、0.01和0.001）、吸附时间（0、30、60、90、120、240 min）、收毒时间（接毒后12、24、36、48、60、72 h）、维持培养温度（35、36、37和38 ℃）和转瓶转速（6、8、10、12、14、16 r/h）7个条件进行优化。结果显示，PEDV CH/GX/750A/2015株在10 L转瓶中的最佳培养条件为：细胞刚铺满单层时接毒，接毒量为MOI=0.01，维持液（无血清DMEM培养液）中胰酶质量浓度为6 μg/mL，不需吸附，维持培养温度为37 ℃，12 r/h 旋转培养48 h后收获病毒液可获得较高效价的病毒液，效价可稳定达到106.50 TCID50/0.1 mL。本试验为 PEDV 变异株大量培养提供了参考，为变异株疫苗研发奠定了基础。
|
| 关 键 词： | 猪流行性腹泻病毒 变异株 转瓶 培养条件 |
Optimization of Large-scale Rotary Bottle Culture Conditions for Porcine Epidemic Diarrhea Virus Mutant CH/GX/750A/2015 |
| |
| LU Bingxia,TAO Minghua,ZHAO Shuo,QIN Yibin,LIU Lei,CHEN Zhongwei,HE Ying,DUAN Qunpeng,LIANG Jiaxing,ZHOU Yingning,LI Bin,JIANG Dongfu,YANG Siyi,ZHAO Wu.Optimization of Large-scale Rotary Bottle Culture Conditions for Porcine Epidemic Diarrhea Virus Mutant CH/GX/750A/2015[J].China Animal Husbandry & Veterinary Medicine,2020,47(5):1506-1513. |
| |
| Authors: | LU Bingxia TAO Minghua ZHAO Shuo QIN Yibin LIU Lei CHEN Zhongwei HE Ying DUAN Qunpeng LIANG Jiaxing ZHOU Yingning LI Bin JIANG Dongfu YANG Siyi ZHAO Wu |
| |
| Institution: | 1. Guangxi Key Laboratory of Veterinary Biotechnology, Guangxi Veterinary Research Institute, Nanning 530001, China;2. Jiangxi Shangrao Animal Husbandry and Veterinary Science Research Institute, Shangrao 334000, China;3. Guangxi University, Nanning 530005, China;4. Guangxi Agricultural Vocational and Technical College, Nanning 530007, China |
| |
| Abstract: | The purpose of this study was to determine the optimal conditions for the rotation culture of CH/GX/750A/2015 strain of porcine epidemic diarrhea virus (PEDV) on Vero cells,and provide technical support for the mass production of high-efficiency vaccine.The virus was cultured on Vero cells in a 10 L spinner flask.The conditions of trypsin concentrations(0,2,4,6,8,10 μg/mL),cell density(40%,60%,80%,100% and 200%),virus inoculation dose (MOI=1,0.1,0.01 and 0.001),virus adsorption time (0,30,60,90,120 and 240 min),virus harvest time (12,24,36,48,60,72 h after virus inoculation),culture temperature after virus inoculation(35,36,37 and 38 ℃) and spinner speed(6,8,10,12,14 and 16 r/h) were optimized.The results showed that the best culture condition of PEDV CH/GX/750A/2015 strain in 10 L transfer flask was to inoculate the virus when the cell was just covered with a single layer,the dose of virus was MOI=0.01,the concentration of trypsin in the culture medium after virus inoculation was 6 μg/mL,without adsorption,the culture temperature after virus inoculation was 37 ℃,the best spinner speed 12 r/h,and the best virus harvest time was 48 h after virus inoculation.After optimization,the virus titer can reach 106.50 TCID50/0.1 mL.This study provided a reference for the large-scale culture of PEDV mutant strain,and laid a foundation for the development of high-efficiency vaccine of PEDV mutant strain. |
| |
| Keywords: | porcine epidemic diarrhea virus variant strain rotary bottle culture condition |
|
| 点击此处可从《中国畜牧兽医》浏览原始摘要信息 |
| 点击此处可从《中国畜牧兽医》下载免费的PDF全文 |
|
