双抗原夹心ELISA检测抗鳗弧菌抗体效价的研究

建立检测抗鳗弧菌抗体效价的双抗原夹心ELISA法,评价其可行性。采用高碘酸盐法对鳗弧菌抗原进行辣根过氧化物酶标记,并通过血清抗体效价比较双抗原夹心ELISA与微量凝集法的优越性。结果显示:标记蛋白含量为2.5mg的抗原所需HRP的最适量为0.20mg。以超声破碎后鳗弧菌离心上清为最适标记抗原,其工作浓度为12.25mg/L,并且双抗原夹心ELISA操作简易具有较高的敏感性和特异性。

In order to develop a specific,sensitive,rapid and practical method that antibodies against Vibrio anguillarum was detected by the improved Double-antigen sandwich ELISA. Vibrio anguillarum is labelled by HRP,and micro-agglutination compares with double-antigen sandwich ELISA by the antibody titer. The results show that labelled antigen which contents 2.5mg protein needs 0.20mg HRP and the centrifugation of Vibrio anguillarum is the optimal labelled antigen togetherwith its working concentration of 12.25mg/L.The established S-ELISA in this study may provide a novel means for detection of Vibrio anguillarum antibody with high sensitivity and specificity.