文章摘要
大口黑鲈弹状病毒的分离培养及其卵黄抗体的制备
The isolation and egg-yolk antibody preparation of Micropterus salmonides rhabdovirus
投稿时间:2019-07-04  修订日期:2019-07-30
DOI:
中文关键词: 大口黑鲈  弹状病毒  卵黄抗体  病毒中和
英文关键词: Micropterus salmoides  rhabdovirus  egg-yolkSantibody  virus neutralization
基金项目:
作者单位E-mail
袁雪梅 农业部淡水渔业健康养殖重点实验室 179506607@qq.com 
吕孙建 农业部淡水渔业健康养殖重点实验室  
施伟达 农业部淡水渔业健康养殖重点实验室  
杭小英 农业部淡水渔业健康养殖重点实验室  
刘莉 农业部淡水渔业健康养殖重点实验室 liuli6655@hotmail.com 
吴颖蕾 农业部淡水渔业健康养殖重点实验室  
摘要点击次数: 282
全文下载次数: 0
中文摘要:
      分离培养大口黑鲈弹状病毒(Micropterus salmoides rhabdovirus,MSRV),并制备卵黄抗体,考察其抗病毒中和作用效果。本文从爆发性死亡的大口黑鲈鱼苗病样中分离培养1株大口黑鲈弹状病毒毒株,采用甲醛灭活大口黑鲈弹状病毒(MSRV),制备佐剂灭活苗,免疫产蛋鸡,收集高免蛋制备MSRV卵黄抗体,测定其效价、中和效果及对病毒复制和宿主细胞内凝集素(intelectin)表达的影响。结果表明,成功利用草鱼卵巢细胞株CO细胞分离培养大口黑鲈鱼弹状病毒,并用于佐剂灭活疫苗的制备;MSRV 佐剂灭活疫苗免疫蛋鸡,成功获得MRSV的卵黄抗体,效价为1:256,稀释度为1/64时的中和病毒的作用最为明显,中和作用率达38.29%以上,病毒核酸拷贝数明显降低。同时还可上调细胞内凝集素的表达。表明特异性卵黄抗体对大口黑鲈弹状病毒具有明显的中和效果,为后续卵黄抗体作为免疫制剂的应用奠定了基础。
英文摘要:
      Largemouth BASS Micropterus salmoides, has been widely cultured in China due to its considerable economic value. Recently, it suffers serious M. salmoides rhabdovirus (MSRV) disease during the juvenile time, particularly in arti?cial aquaculture. In this study, a strain of Micropterus salmoides rhabdovirus was isolated from dying juvenile fish, and cultured in CO cell line. Considering the advantage of passive immunity directly against MSRV, we explored the anti-MSRV IgY by discontinuously injecting the formalin-killed vaccine of MSRV into the laying hens. The antibody was purified by saturated ammonium sulphate, and displayed virus resistance with ElISA antibody titer of 1:256 detected. The bioactivity of anti-MSRV IgY was tested simultaneously after being added into the grass carp ovary cell (CO) line incubated with MSRV. This antibody showed antiviral activity with 38.29% of neutralizing effectiveness detected at the dilution of 1:64. The RNA copies of MSRV in test group were obvirously decreased compared with control group. By the florescent real-time quantitative RT-PCR, the intracellular lectin experienced significantly up-regulated and pick up to nearly 10 folds at 72 h as compared with control group. In conclusion, we succeeded to isolated a strain of MRSV and cultured in CO cell line. Meanwhile, anti-MSRV IgY was prepared with vaccinations of inactive virus and displayed antiviral activity in this study, which laid a foundation for the application of yolk antibody as immune preparation.
相关附件:   附件1  附件1  附件2
View Fulltext   查看/发表评论  下载PDF阅读器
关闭