鱼肠道弧菌免疫检测方法的研究  

Studies on Immunological Detection Methods for Vibrio ichthyoenteri

作　　者：刘慧[1] 邓灯[1] 王忠[1] 程顺峰[1,2] 孙玉霞[3] 

机构地区：[1]青岛农业大学动物科技学院,山东青岛266109 [2]淮海工学院江苏省海洋生物技术重点建设实验室 [3]山东省农业科学院

出　　处：《青岛农业大学学报（自然科学版）》2011年第3期223-226,共4页Journal of Qingdao Agricultural University(Natural Science)

基　　金：国家自然科学基金项目(30901106);山东省自然科学基金项目(ZR2009DM036);淮海工学院江苏省海洋生物技术重点建设实验室研究基金(2009HS11);青岛农业大学高层次人才科研基金(630701)资助

摘　　要：采用灭活的鱼肠道弧菌作为抗原,制备兔抗血清,建立了鱼肠道弧菌的间接荧光抗体检测技术(indirect im-munofluorescence assay test,IFAT)、Western-blotting免疫印迹检测技术。IFAT结果显示阳性信号覆盖整个菌体,明亮,清晰,荧光信号呈长弧形且两端钝圆,与鱼肠道弧菌形状一致。Western-blotting结果显示共10条蛋白带发生免疫反应,其中6条蛋白带结果较清晰,显示出较强的特异性免疫反应,阴性对照组无条带。IFAT和Western-blotting结果说明所建立检测方法能够准确、快速的检测出鱼肠道弧菌。In this paper, indirect immunofluorescence assay test（IFAT）and Western -blotting for rapid diagnosis of Vibrio ichthyoenteri have been developed. Antiserum specific for the Vibrio ichthyoenteri was produced from New Zealand white rabbit and used in IFAT and Western - blotting. The consequent of IFAT showed that light and clear positive signals covered the whole bacteria with the shape of long arc and roundness which fitted with Vibrio ichthyoenteri. Western blotting analysis revealed ten antigenic protein bands. Six major antigens with apparent molecular masses of approximately 88, 79, 72, 61,42 and 18 kDa as well as other four less immunoractive bands of approximately102, 36, 34 and 29 kDa were distinguished. This study will facilitate the development of rapid diag- nostic techniques for Vibrio ichthyoenteri.

关 键 词：鱼肠道弧菌 间接荧光抗体技术 免疫印迹技术 检测技术 

分 类 号：S941.42[农业科学—水产养殖]