| 鲤疱疹病毒2型(CyHV-2)灭活疫苗制备及其免疫效力测定 |
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| 引用本文： | 潘晓艺,蔺凌云,朱悦,姚嘉赟,尹文林,刘忆瀚,陈凡,沈锦玉.鲤疱疹病毒2型(CyHV-2)灭活疫苗制备及其免疫效力测定[J].海洋与湖沼,2022,53(5):1143-1150. |
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| 作者姓名： | 潘晓艺 蔺凌云 朱悦 姚嘉赟 尹文林 刘忆瀚 陈凡 沈锦玉 |
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| 作者单位： | 浙江省淡水水产研究所 农业农村部淡水渔业健康养殖重点实验室 浙江省鱼类健康与营养重点实验室 浙江湖州 313001;浙江海洋大学水产学院 浙江舟山 316022;杭州市农业技术推广中心 浙江杭州 310001 |
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| 基金项目： | 浙江省重点研发计划,2019C02058号;浙江省科研院所扶持专项,2021YSZX001号。 |
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| 摘 要： | 鲫造血器官坏死病是由鲤疱疹病毒2型(CyHV-2)引起的危害我国养殖鲫鱼(Carassius auratus)最严重的病毒性疾病。为制备有效预防鲫造血器官坏死病的灭活疫苗,以CyHV-2代表流行株CNDF-TB2015为疫苗株,以CSC细胞系进行病毒增殖,并采用不同浓度的病毒对不同规格鲫鱼进行攻毒,建立攻毒模型。结果表明,培养的病毒滴度达1010 TCID50/mL,以4×107TCID50/尾的剂量腹腔注射感染体重300 g的鲫鱼可造成100%的死亡率。对病毒灭活剂甲醛、β-丙内酯和BEI灭活条件进行筛选优化,对疫苗免疫佐剂763A、IMS1312、铝胶和M402进行筛选,结果表明,浓度为108.3 TCID50/mL的病毒液最佳灭活条件为终浓度10 mmol/L的BEI 37 ℃灭活72 h,疫苗制备最佳佐剂为水佐剂IMS1312。对灭活疫苗的最小免疫剂量、最佳免疫剂量和有效免疫产生期进行测定,结果表明,要获得70%以上保护率的最小免疫剂量为8×106 TCID50/尾,最佳免疫剂量为4×107 TCID50/尾,在免疫4周后保护率可达80%, 5周后保护率达到100%。制备的灭活疫苗为鲫造血器官坏死病的免疫预防提供了有力手段,也为其他水产动物病毒灭活疫苗的制备提供了借鉴。
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| 关 键 词： | 鲤疱疹病毒2型 鲫造血器官坏死病 灭活疫苗 免疫效力 |
| 收稿时间： | 2021/12/16 0:00:00 |
| 修稿时间： | 2022/1/22 0:00:00 |
PREPARATION AND EFFICACY OF INACTIVATED VACCINE AGAINST CYPRINID HERPESVIRUS-2 |
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| PAN Xiao-Yi,LIN Ling-Yun,ZHU Yue,YAO Jia-Yun,YIN Wen-Lin,LIU Yi-Han,CHEN Fan,SHEN Jin-Yu.PREPARATION AND EFFICACY OF INACTIVATED VACCINE AGAINST CYPRINID HERPESVIRUS-2[J].Oceanologia Et Limnologia Sinica,2022,53(5):1143-1150. |
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| Authors: | PAN Xiao-Yi LIN Ling-Yun ZHU Yue YAO Jia-Yun YIN Wen-Lin LIU Yi-Han CHEN Fan SHEN Jin-Yu |
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| Institution: | Ministry of Agriculture and rural areas Key Laboratory of Healthy Freshwater Aquaculture, Key Laboratory of Fish Health and Nutrition of Zhejiang Province, Zhejiang Institute of Freshwater Fisheries, Huzhou 313001, China;School of Fishery, Zhejiang Ocean University, Zhoushan 316022, China;Hangzhou Centre for Agricultural Technology Extension, Hangzhou 310001, China |
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| Abstract: | Herpesviral haematopoietic necrosis disease (HVHN) is a viral disease of crucian carp, caused by Cyprinid herpesvirus-2 (CyHV-2), which causes huge losses on aquaculture in China. In order to control the occurrence of HVHN, the inactivated vaccine was developed based on the vaccine strain CNDF-TB2015 and the immune efficacy was assessed. The results of CyHV-2 culture showed that the titer of virus cultured with CSC cells reached 1010 TCID50/mL, and the challenge model test showed the dose of 4×107 TCID50/ind could cause 100% mortality in crucian carp within 14 d post injection with the CyHV-2 CNDF-TB2015. The results of inactivated vaccine preparation test showed that the optimal condition for CyHV-2 inactivation with 108.3 TCID50/mL was 10 mM BEI at 37 °C for 72 h and the best adjuvant was IMS1312. The protection rate with the minimum immune dose of 8×106 TCID50 /ind could reach more than 70%, with the optimal immune dose of 4×107 TCID50/ind could reach 100%. The protection rate reached 80% at week 4 postimmune and 100% at week 5 postimmune. This study indicated that the inactivated CNDF-TB2015 strain of CyHV-2 could serve as a potential vaccine candidate to prevent the prevalence of HVHN, also provides reference for the preparation of other inactivated viral vaccines for aquaculture. |
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| Keywords: | CyHV-2 HVHN inactivated vaccine immune efficacy |
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