Dot-ELISA快速检测鱼类运动性气单胞菌的研究

Rapid Detection of Motile Aeromonas Species in Fish by Dot ELISA

制备了引起常见鱼病的６株运动性气单胞菌的兔抗血清，建立了检测运动性气单胞菌的Ｄｏｔ－ＥＬＩＳＡ法。该法能检出含１０７菌细胞／ｍＬ的菌悬液及１０个菌细胞／ｍＬ增菌２０ｈ的培养物，与运动性气单胞菌群内的菌株，均呈现明显的阳性反应，不与鲁克耶尔森氏菌、荧光假单胞菌、鳗弧菌等１０株对照菌发生影响检测结果的交叉反应。应用Ｄｏｔ－ＥＬＩＳＡ与常规分离法分别对人工感染的４０份样本进行检测，其阳性数分别为３３和３４，两法符合率为９７％。经Ｘ２检验表明，两法检测结果间有显著意义的一致性（Ｐ＜０．０５），但前者可在２４ｈ内报告结果。应用Ｄｏｔ－ＥＬＩＳＡ又对８株鱼源菌检定，结果与细菌学检查相符。Ｄｏｔ－ＥＬＩＳＡ法具有快速、敏感、特异、实用的优点，可用于引起常见鱼病的运动性气单胞菌群的检测。

Dot ELISA was developed to detect 6 strains of motile Aeromonas species from fish. It is able to detect 10 7 cells/mL in the bacteium suspension or 10 cells/mL of culture enriched for 20 h, and no cross reactions occured with 10 other species of bacteria including Yersinia ruckeri, Pseudomonas fluorescens and Vibril anguillarum , etc. Of 40 artificial specimens, 33 and 34 were detected by Dot ELISA and by the conventional culture technique, respectively. The consistence between the results of the two methods were statisticlly significant (p<0.05) (the consistent rate of 97.5%). The results can be reported in 24 hrs by the Dot ELISA. The results of the Dot ELISA confirm the results of bacteriological examination in identifying 8 strains of bactoria from fishes. The Dot ELISA is characterized by specificity, sensitivity, time saving and feasibility and can be used in a laboratory.