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以梭子蟹乳化病的病原菌-溶藻弧菌为抗原,制成免疫血清;利用辣根过氧化酶标记的羊抗兔血清(HRP-IgG)为酶标二抗、建立了检测溶藻弧菌的间接ELISA快速诊断法。结果表明,应用间接ELISA技术检测溶藻弧菌有较高的灵敏度,最低的检测量为104cfu/mL。同时交叉反应表明其具有较高的特异性。人工感染试验48 h后,就能在血淋巴和鳃中检测到溶藻弧菌。应用建立的间接ELISA法进行现场检测,表观有病蟹的阳性检出率为90%,表观健康蟹的阳性检出率为20%,养殖海水中没有阳性检出。
The immune serum was made by using Vibrio alginolyticus as the pathogen of emulsifying disease of shuttle crabs. HRP-IgG labeled with horseradish peroxidase (HRP-IgG) was used as secondary antibody to detect Vibrio alginolyticus Indirect ELISA rapid diagnostic method. The results showed that the application of indirect ELISA technology to detect V. alginolyticus has a higher sensitivity, the lowest detection of 104cfu / mL. At the same time cross-reaction showed that it has high specificity. 48 hours after the artificial infection test, V. alginolyticus could be detected in the hemolymph and gills. Indirect ELISA method was used to detect the spot, the positive detection rate of sick crab was 90%, the positive detection rate of healthy crab was 20%, there was no positive detection in cultured seawater.