鲤疱疹病毒2型(CyHV-2)灭活疫苗制备及其免疫效力测定  

PREPARATION AND EFFICACY OF INACTIVATED VACCINE AGAINST CYPRINID HERPESVIRUS-2

作　　者：潘晓艺[1] 蔺凌云[1] 朱悦 姚嘉赟[1] 尹文林[1] 刘忆瀚 陈凡 沈锦玉[1,2] PAN Xiao-Yi;LIN Ling-Yun;ZHU Yue;YAO Jia-Yun;YIN Wen-Lin;LIU Yi-Han;CHEN Fan;SHEN Jin-Yu(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)

机构地区：[1]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 [2]浙江海洋大学水产学院,浙江舟山316022 [3]杭州市农业技术推广中心,浙江杭州310001

出　　处：《海洋与湖沼》2022年第5期1143-1150,共8页Oceanologia Et Limnologia Sinica

基　　金：浙江省重点研发计划,2019C02058号;浙江省科研院所扶持专项,2021YSZX001号。

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10^(10)TCID_(50)/mL,以4×10^(7)TCID_(50)/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10^(8.3) TCID_(50)/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8×10^(6)TCID_(50)/尾,最佳免疫剂量为4×10^(7)TCID_(50)/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。Herpesviral haematopoietic necrosis(HVHN)is a viral disease of crucian carp caused by Cyprinid herpesvirus-2(CyHV-2),which causes huge losses on aquaculture in China.To prevent from the outbreak of HVHN,the inactivated vaccine was developed using vaccine strain CNDF-TB2015 and the immune efficacy was assessed.The CyHV-2 culture showed that the titer of virus cultured with CSC cells reached 10^(10)TCID_(50)/mL,and the challenge model test showed the dose of 4×10^(7)TCID_(50)/ind could cause 100%mortality in crucian carp within 14 d post injection with the CyHV-2 CNDF-TB2015.The inactivated vaccine preparation test showed that the optimal inactivation condition for CyHV-2 at 10^(8.3) TCID_(50)/mL was 10 mmol/L BEI at 37℃ for 72 h and the best adjuvant was IMS1312.The minimum immune dose was 8×10^(6)TCID_(50)/ind,and the optimal immune dose was 4×10^(7)TCID_(50)/ind.The protection rate reached 80%in 4 weeks and 100%in 5 weeks after immunization with dose of 4×10^(7)TCID_(50)/ind.This study presents that the inactivated CNDF-TB2015 strain of CyHV-2 could serve as a potential vaccine candidate to prevent the prevalence of HVHN,and provides a reference for the preparation of other inactivated viral vaccines for fish in aquaculture.

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 

分 类 号：S942.5[农业科学—水产养殖]