| 基于CPB细胞扩增体系的鳜弹状病毒增殖条件的优化 |
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| 引用本文： | 罗霞,付小哲,李宁求,林强,黄志斌,张悠. 基于CPB细胞扩增体系的鳜弹状病毒增殖条件的优化[J]. 水产学报, 2018, 42(9): 1481-1488 |
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| 作者姓名： | 罗霞 付小哲 李宁求 林强 黄志斌 张悠 |
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| 作者单位： | 中国水产科学研究院珠江水产研究所农业部渔用药物创制重点实验室广东省水产动物免疫技术重点实验室 |
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| 基金项目： | 广东省渔港建设和渔业产业发展专项（粤财农〔2017〕17号）；广东省省级科技计划项目（2016B020234003）；广东省省级科技计划项目（2016A020210021） |
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| 摘 要： | 为获知鳜弹状病毒(Siniperca chuatsi rhabdovirus,SCRV)QY株在体外培养细胞中最适增殖条件,以鳜脑细胞系(Chinese perch brain cell line,CPB)为增殖体系,采用实时荧光定量PCR(quantitative real-time PCR,q RT-PCR)技术所测病毒拷贝数为判断指标,比较同步接毒和异步接毒2种接种方式以及病毒接种量、血清浓度等培养条件对病毒增殖的影响,确定SCRV-QY株在CPB细胞中的最适增殖条件。结果显示,单位体积病毒增殖量方面,异步接毒法优于同步接毒法,以感染复数(multiplicity of infection,MOI)为10将病毒接种长满单层的CPB细胞中,28°C吸附1.5 h,用含2%胎牛血清的L15培养液于28°C培养时,病毒增殖量最高,为5.59×10~(10)拷贝/mL;而从单位成本所获病毒量考虑,同步接毒法筛选出的4种增殖条件单位成本所获的病毒产量优于异步接毒法,其中当MOI=0.03、胎牛血清终浓度为6%时,同步接种对数生长中期的CPB细胞,28°C恒温培养70 h后收获病毒液,单位成本所获病毒量最高,为4.88×10~(13)个拷贝/元。综上所述,本研究以病毒增殖量和培养基成本作为考量,优化SCRV-QY株的增殖条件,可为SCRV疫苗低成本、规模化生产提供理论依据。
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| 关 键 词： | 鳜弹状病毒 鳜脑细胞 接毒方法 增殖条件 实时荧光定量PCR |
| 收稿时间： | 2017-06-05 |
| 修稿时间： | 2017-08-30 |
Study on optimal proliferation condition of Siniperca chuatsi rhabdovirus in CPB cells |
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| LUO Xi,FU Xiaozhe,LI Ningqiu,LIN Qiang,HUANG Zhibin and ZHANG You. Study on optimal proliferation condition of Siniperca chuatsi rhabdovirus in CPB cells[J]. Journal of Fisheries of China, 2018, 42(9): 1481-1488 |
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| Authors: | LUO Xi FU Xiaozhe LI Ningqiu LIN Qiang HUANG Zhibin ZHANG You |
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| Affiliation: | Key Laboratory of Fishery Drug Development, Ministry of Agriculture;Key Laboratory of Aquatic Animal Immune Technology, Guangdong Province Pearl River Fishery Research Institute, Chinese Academy of Fishery Sciences, Guangzhou 510380, China,Key Laboratory of Fishery Drug Development, Ministry of Agriculture;Key Laboratory of Aquatic Animal Immune Technology, Guangdong Province Pearl River Fishery Research Institute, Chinese Academy of Fishery Sciences, Guangzhou 510380, China,Key Laboratory of Fishery Drug Development, Ministry of Agriculture;Key Laboratory of Aquatic Animal Immune Technology, Guangdong Province Pearl River Fishery Research Institute, Chinese Academy of Fishery Sciences, Guangzhou 510380, China,Key Laboratory of Fishery Drug Development, Ministry of Agriculture;Key Laboratory of Aquatic Animal Immune Technology, Guangdong Province Pearl River Fishery Research Institute, Chinese Academy of Fishery Sciences, Guangzhou 510380, China,Key Laboratory of Fishery Drug Development, Ministry of Agriculture;Key Laboratory of Aquatic Animal Immune Technology, Guangdong Province Pearl River Fishery Research Institute, Chinese Academy of Fishery Sciences, Guangzhou 510380, China and Key Laboratory of Fishery Drug Development, Ministry of Agriculture;Key Laboratory of Aquatic Animal Immune Technology, Guangdong Province Pearl River Fishery Research Institute, Chinese Academy of Fishery Sciences, Guangzhou 510380, China |
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| Abstract: | |
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| Keywords: | Siniperca chuatsi rhabdovirus (SCRV) Chinese perch brain cell line, CPB cells inoculation method proliferation condition quantitative real-time PCR |
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