| In this research, Aeromonas hydrophila YT-1 were isolated from the diseased eel with septicemia in Fujian. The results of the hemolysis assay , PCR and the virulence assay demonstrated that the bacterial YT-1 could lead to eel and mice death. Furthermore, the bacterial could produce an extracellular toxin which possessed hemolytic activity and lethal virulence. Monoclonal antibodies against the bacterial YT-1 and AH10501 (the standard strain) were prepared by hybridizing the Sp2/0 myeloma cell with the spleen cell from immunized mice. Eleven hybridoma cell lines secreting monoclonal antibodies(McAbs) against Aeromonas hydrophila had been selected from the antibody positive hybridizing cells. They were named as AF11.FA4, AG4, CA4, EE8, CF2, DCS, GH9, 2F9, 2H4 and 4G4 respectively. Isotyping anal ysis demanstratcd that these McAbs belong to IgM or IgG2 subclasses. The 1-XISA titres of these McAbs were ranged between 10-4?0-6. And McAb 2F9 had agglutination ability with three isolates of Aeromonas hydrophila . Specificity analysis proved that McAb 4G4 and 2H4 only recognized the Aeromonas hydrophila isolates belong to 0: AH10501 serotype, and the McAb FA4, GH9 and CF2 reacted with the bacterials belong to 0: YT-1 serotype, and all of them did not have any cross reaction with the other Aeromonas, Vibrio anguillarum, Edwardsiella^ Ycrsinia ruckcri, Salmonella, Eschcrichia coli, Pseudomonas fluorescens and Pseudomonas putrefaciens. Western-blotting assay demonstrated that McAb 404,2114 and FA4 bound to the different lipopolysaccharide (LPS) fraction of Aeromonas hydrophila. And also McAb GH9, 2H4 and FA4 were tested successfully for their ability to be used for thedetection of the homologous serotype strains by immunofluorescence microscopy. All results indicated that McAb 4G<^ 2H4 and GH9 had the great potential to be applied to clinical diagnosis and identify the serotype of Aeromonas hydrophila.
|
PDF Full Text Request