| 抗鲤疱疹Ⅱ型病毒卵黄抗体制备及其功能鉴定 |
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| 引用本文： | 寇海燕,郭培红,田丹阳,何稳,李坤,张婷婷,贡成良,孙丙耀,宋学宏.抗鲤疱疹Ⅱ型病毒卵黄抗体制备及其功能鉴定[J].淡水渔业,2021,51(1):65-71. |
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| 作者姓名： | 寇海燕 郭培红 田丹阳 何稳 李坤 张婷婷 贡成良 孙丙耀 宋学宏 |
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| 作者单位： | 苏州大学医学部基础医学与生物科学学院,江苏苏州215123;昆山贝瑞康生物科技有限公司,江苏昆山215345 |
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| 基金项目： | 江苏省渔业科技类项目(Y2018-19)。 |
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| 摘 要： | 为制备抗鲤疱疹Ⅱ型病毒(CyHV-2)的卵黄抗体,探索防治异育银鲫(Carassius auratus gibelio)鳃出血病的新方法和途径,本研究利用原核表达系统产生具免疫原性的重组CyHV-2-ORF72衣壳蛋白,纯化后免疫蛋鸡;二次免疫后采用间接ELISA法抽检免疫蛋的特异性卵黄抗体(IgY)含量,收集抗体效价...
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| 关 键 词： | 卵黄抗体 鲤疱疹Ⅱ型病毒 异育银鲫(Carassius auratus gibelio) 免疫保护 |
Preparation and functional identification of egg yolk antibody against cyprinid herpesvirus 2 |
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| KOU Hai-yan,GUO Pei-hong,TIAN Dan-yang,HE Wen,LI Kun,ZHANG Ting-ting,GONG Cheng-liang,SUN Bing-yao,SONG Xue-hong.Preparation and functional identification of egg yolk antibody against cyprinid herpesvirus 2[J].Freshwater Fisheries,2021,51(1):65-71. |
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| Authors: | KOU Hai-yan GUO Pei-hong TIAN Dan-yang HE Wen LI Kun ZHANG Ting-ting GONG Cheng-liang SUN Bing-yao SONG Xue-hong |
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| Institution: | (School of Biology and Basic Medical Sciences,Soochow University,Suzhou 215123,Jiangsu,China;Kunshan Best-Care Biological Technology Co.Ltd.,Kunshan 215345,Jiangsu,China) |
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| Abstract: | This experiment generated recombinant CyHV-2-ORF72 capsid protein with potential immunogenicity in prokaryotic expression system to produce egg yolk antibody against cyprinid herpesvirus 2(CyHV-2)and explore new methods and approaches to prevent and treat haemorrhagic gill disease of allogynogenetic crucian carp.The recombinant ORF72 protein was purified and used to immunize the laying hens.After the second immunization,random sampling was performed to determine the contents of specific yolk antibody(IgY)in immunized eggs by indirect ELISA.The immunized eggs with antibody titer reaching 1:8000 were harvested and divided into whole egg group and egg yolk group,and then the whole egg powder and egg yolk powder were prepared by freeze-drying technology after micro-encapsulation.The specificity and titer of IgY in the whole egg powder and egg yolk powder were then detected by using Western blot and ELISA techniques.Next,in vitro neutralization test was performed by incubating the CyHV-2-infected epithelioma papulosum cyprini cells with egg yolk powder diluents.In the full-scale therapeutic trial,the available feed was prepared by incorporating the whole egg powder with anti-CyHV-2-ORF72 antibody into a commercial feed with the addition of 0.15%and 0.30%,respectively,and fed to the sick fish in two ponds.The results showed that IgY in both whole egg powder and egg yolk powder could specifically bind to CyHV-2-ORF72 capsid protein,with titers ranging from 1:8000 to 1:16000.The anti-CyHV-2-ORF72 IgY had a high affinity with CyHV-2,and its neutralizing activity was higher than that of the mouse anti-ORF72 serum.Moreover,74.3%and 87.6%fish in the two experimental ponds survived,while only 28.3%and 31.5%fish in control ponds survived.Collectively,the anti-CyHV-2 egg yolk antibody prepared in this study shows the characteristics of virus neutralization and exhibits adequate immunoprotection against CyHV-2. |
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| Keywords: | egg yolk immunoglobulin cyprinid herpesvirus 2 allogynogenetic crucian carp immune protection |
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