Detection of Vibrio anguillarum and Its Virulent Metalloprotease Using the Method of ELISA

A method detecting pathogenic Vibrio anguillarum and its virulent metalloprotease is reported in this paper. The metalloprotease is isolated from extracellular product (ECP) of V.anguillarum by the cellophane plate technique and purified by gel filtration and ion-exchange chromatography. Anti-sera are prepared by injecting V.anguillarum cells and metalloprotease into the rabbits. Slide Agglutination Assay is used to detect V.anguillarum in the infection experiment and Enzyme Linked Immunosorbent Assay (ELISA) is carried out to detect concentration of metalloprotease. The result shows that bacterium strain M3 is able to diffuse into the viscera of infected fish through the blood circulating system 10 hours after intramuscular infection, and ELASA is a sensitive method to detect the metalloprotease with detectable amount of 7.8ng. The aim of this study is to establish a sensitive and specific method to observe the infection of V.anguillarum in the host.