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采用菌体吸附法对海豚链球菌(Streptococcus iniae)、迟缓爱德华氏菌(Edwardsiella tarda)、鳗弧菌(Vibrio anguillarum)、杀鲑气单胞菌(Aeromonas salmonicida)、荧光假单胞菌(Psedomonas fluo-rescens)和海洋分支杆菌(Mycobacterium marinum)6种养殖鱼类病原菌的兔抗血清进行吸附,以减小抗血清与其它菌株的交叉反应;吸附纯化后的抗体作为捕获抗体构建了病原菌检测抗体芯片。实验结果显示,与未经过吸附的纯化抗血清构建的抗体芯片相比,血清吸附实验能有效减少交叉反应,降低抗体的非特异性吸附,提高细菌检测的特异性。同时,以迟缓爱德华氏菌为例,采用吸附纯化后抗体制备的抗体芯片检测人工感染迟缓爱德华氏菌和自然发病的牙鲆,均观察到迟缓爱德华氏菌的特异性显色反应,得到了较理想的检测结果。本文结果说明优化后的病原菌检测抗体芯片可用于水产养殖鱼类常见的上述6种细菌性疾病的有效检测。
Streptococcus iniae, Edwardsiella tarda, Vibrio anguillarum, Aeromonas salmonicida, Psedomonas fluo -rescens) and Mycobacterium marinum were used to reduce the cross-reaction between antiserum and other strains. The purified antibody was used as the capture antibody to construct the pathogen detection antibody chip . The experimental results show that, compared with the antibody chip constructed without purification of the purified antiserum, the serum adsorption experiment can effectively reduce the cross-reaction, reduce the nonspecific adsorption of the antibody, and improve the specificity of the bacterial detection. At the same time, taking Edwardsiella tarda as an example, the specific color reaction of Edwardsiella tarda was observed by using antibody chip prepared by adsorption purification of purified antibodies against Edwardsiella spp. The ideal test result. The results of this study demonstrate that the optimized pathogen detection antibody chip can be used for the effective detection of the above six bacterial diseases common to aquaculture fish.