草鱼细菌性烂鳃病病原菌的分离及其特异性卵黄抗体的初步研究  

Study on IgY Against the Pathogen of the Bacterial Gill Rot Disease in Grass Carp

作　　者：李思泉 李娜 罗玉双 张保平[3] 张建平 Li Siquan;Li Na;Luo Yushuang;Zhang Baoping;Zhang Jianping(Life and Environment Sciences College of Hunan University of Arts and Science,Changde Hunan 415000,China;Key Laboratory of Health Aquaculture and Product Processing in Dongting Lake Area of Hunan Province,Zoology Key Laboratory of Hunan Higher Education,Changde Hunan 415000,China;Changde Vocational Technical College,Department of Agriculture and Forestry Engineering,Changde Hunan 415000,China)

机构地区：[1]湖南文理学院生命与环境科学学院,湖南常德415000 [2]环洞庭湖水产健康养殖及加工湖南省重点实验室,动物学湖南省高校重点实验室,湖南常德415000 [3]常德职业技术学院农林工程系,湖南常德415000

出　　处：《贵州畜牧兽医》2019年第3期37-42,共6页Guizhou Journal of Animal Husbandry & Veterinary Medicine

基　　金：“草鱼柱状黄杆菌卵黄抗体IgY的研制及其抑菌功能的初步评价”湖南省教育厅科学研究项目(14C0805);“鸡抗嗜水气单胞菌高免疫卵黄抗体的制备”湖南文理学院科研项目(16YB18);“抗黄杆菌卵黄抗体的制备及其被动免疫保护作用的研究”湖南文理学院生命科学学院省级平台开放项目(SKYPT201607)

摘　　要：试验从患典型烂鳃病草鱼病变部位分离到1株柱状黄杆菌(FC-3株),采用0.4%甲醛灭活制成免疫原,接种健康120日龄桃源蛋鸡,收集鸡蛋并提纯卵黄抗体(IgY),并以此IgY为基础建立间接ELISA方法。结果:用FC-3株制备的IgY经初步提纯后电泳分离可见重链和轻链2条蛋白条带,获得了IgY多克隆抗体。建立的间接ELISA方法检测柱状黄杆菌纯培养液的检出限值为1×10^7cfu/mL,抗原包被最佳浓度和抗体最佳工作浓度分别为1∶100和1∶512,辣根过氧化物酶标记兔抗鸡IgY最佳工作浓度为1∶4000。结论:建立的间接ELISA方法对柱状黄杆菌有明显的检测特异性,与其他菌株无交叉反应,重复性好。Streak plate method was used to separate the pathogenic strain FC-3 from diseased regions of Ctenopharyngodon idella with gill rot,named Flavobacterium columnare.The strain was inactivated by 0.4%formaldehyde according to the immune plan to stimulate 120 days laying Taoyuan hens’the production of Yolk antibody(IgY).The high level immuned eggs were collected.The purified IgY was used to establish indirect ELISA for detecting the FC-3 strain.Results:the Flavobacterium columnare(FC-3)IgY was preliminary purified,2 protein strips heavy chain and light chain were visible by Electrophoresis separation.This indicate the IgY polyclonal antibody was acquired.Titer of the polystyrene chloride microtitration plates were coated with Flavobacterium columnare antigen were detected by IgY indirect ELISA at the amount of 1×10 7 cfu/mL.The working concentration of antigen,primary antibody and HRP-labelled rabbit antichicken IgY were 1∶100,1∶512 and 1∶4 000.Conclusion:the detection specificity of the method was also determined,Flavobacterium columnare can be detected,while other bacteria strains can not be detected,good repeatability.

关 键 词：柱状黄杆菌 卵黄抗体 间接ELISA 

分 类 号：S941.42[农业科学—水产养殖]