Establishment and application of biotin-avidin-amplified enzyme-linked immunosorbent assay on Aeromonas hydrophila of Chinese soft-shelled turtles, Pelodiscus sinensis
2008
Hu Dayan | Qian Dong | Liu Wen
Chinese. 从中华鳖中分离的嗜水气单胞菌致病株T0608经福尔马林灭活后,注射免疫新西兰兔,制备了高价免疫血清,血清的菌体凝集效价可达1/1 280;G蛋白亲和层析纯化兔IgG,将IgG用生物素标记,建立了生物素-亲和素双抗体夹心酶联免疫吸附法(BA-ELISA);其最低检出限为2.5×103cfu,相当于0.5 ng菌体蛋白,其检测灵敏度为ABC-ELISA(4.0×104cfu)和间接ELISA(4.0×104cfu)的16倍;特异性试验结果表明,本法与鱼类气单胞菌、拟态弧菌、海豚链球菌、大肠杆菌等无交叉反应.BA-ELISA可不经细菌培养直接检测人工感染48 h中华鳖的肝、肠道组织中的嗜水气单胞菌.用该方法对湖州市龟鳖养殖场的水样、中华鳖和巴西龟肝、肠道内容物进行检测.结果表明该方法可较好地应用于发病中华鳖的病原早期诊断及养殖中华鳖的病原检测.
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