抗鲶爱德华氏菌多克隆抗体的制备及特性分析

Production and Characterization of Polyclonal Antibodies Against Edwardsiella ictaluri

利用2株黄颡鱼"红头病"病原菌——鲶爱德华氏菌的灭活疫苗作为免疫原,免疫新西兰大白兔,制备了高效价的多克隆抗体。通过间接酶联免疫、间接免疫荧光和免疫印迹等技术对多克隆抗体特性进行了分析。测定获得多抗效价分别为1∶215和1∶214,2株病原菌之间存在很强的交叉反应,且2种多抗均与鲶爱德华氏菌、迟钝爱德华氏菌、副溶血弧菌、鳗弧菌、溶藻弧菌、火神弧菌、河流弧菌、创伤弧菌标准菌株存在交叉反应,与粪肠球菌、大肠杆菌、酿脓链球菌、海豚链球菌、杀鲑气单胞菌、嗜水气单胞菌、类产碱假单胞菌标准菌株均无交叉反应。免疫印迹实验结果表明,2株病原菌的主要抗原决定簇相同,分子量分别为61.1,46.6,41.3,28.8和19 kDa。本实验得到了黄颡鱼"红头病"致病菌高效价的多克隆抗体,可以作为初步筛选"红头病"病原菌的工具,为建立快速有效的疾病监测方法和进一步的免疫学防病研究提供了依据,奠定了基础。

High titer polyclonal antibodies （PAb） against A51 and A97, pathogen of red-head disease in yellow catfish Pelteobagrus fulvidraco, were produced from New Zealand white rabbit with the inactivated A51 and A97. The characterizations of the polyclonal antibodies were analyzed with indirect enzymelined immunosorbent assay, immunofluorescence assay and western blot. The results showed that the titer of the polyclonal antibodies were 1 ： 21^15and 1 ： 2^14 , respectively. There was strong cross-reaction between A51 and A97, and both the PAbs of A51 and A97 could recognize some other strains, such as V. parahaemolyticus, V. anguillarum, V. alginolyticus, V. logei, V. fluvialis, V. vulnificus, but no f cross withE, faecalis, E. coli, S. pyrogenes, S. iniae, A. salmonicida, A. hydrophila, Ps. pseudoalcaligene. Under reducing conditions in western blotting, the major antigenic determinants of the pathogens A51 and A97 were identical, MW was 61.1 kDa, 46.6 kDa, 41.3 kDa, 28. 8 kDa and 19 kDa, respectively. The two PAbs could be used as probe for preliminary screen of pathogen of red-head disease in yellow catfish Pelteobagrus fulvidraco.