Article ID Journal Published Year Pages File Type
2428579 Comparative Immunology, Microbiology and Infectious Diseases 2010 12 Pages PDF
Abstract

Edwardsiella tarda is an important cause for hemorrhagic septicemia in fish and gastro and extra-intestinal infections in humans. Monoclonal antibodies (MAbs) were produced against outer membrane proteins (OMPs) of E. tarda ET-7, isolated from diseased snakehead (Ophiocephalus punctatus). Two stable hybridoma clones, designated as 3F10 and 2C3 MAbs were found to be potentially specific for E. tarda by indirect enzyme linked immunosorbent assay (ELISA). These MAbs recognized major immunogenic OMP band at 44 kDa in Western blotting. Both MAbs belonged to the IgG1 isotype and recognized different epitopes of OMP as seen by competitive ELISA. These MAbs strongly reacted with all 17 isolates of E. tarda used in our study by indirect ELISA and Western blotting. Interestingly, no reaction was observed with the reference strain of E. tarda (MTCC 2400). The sensitivity of 3F10 MAb to detect whole cells of E. tarda was up to a level of 1 × 104 CFU/ml in indirect ELISA. No cross-reactivity of MAbs were seen with Escherichia coli, Salmonella arizonae, Pseudomonas fluorescens, Aeromonas hydrophila, Vibrio cholerae, Flavobacterium ferrugineum and Mycobacterium tuberculosis. These MAbs could be used for specific detection of E. tarda infection in fish by immunoassays.

RésuméEdwardsiella tarda est une cause importante de la septicémie hémorragique chez les poissons et pour les infections gastro-intestinales et extra-intestinales de l’homme. Des anticorps monoclonaux (MAbs) ont été produits contre les protéines externes de membrane (OMPs) de E. tarda ET-7, isolée à partir de Ophiocephalus punctatus. Deux clones d’hybridome désignés comme 3F10 et 2C3 MAbs se sont avérés spécifiques de la détection du E. tarda par ELISA. Ces MAbs ont permis de reconnaître les OMPS de 44 kDa et ont réagi fortement avec tous les isolats de E. tarda utilisés à partir de poissons et d’eau d’étang (17). Il n’y a pas eu d’activité hétérospécifique avec des anticorps monoclonaux ou des cellules bactériennes entières de Escherichia coli, Salmonella arizonae, Pseudomonas fluorescens, Aeromonas hydrophila, Vibrio cholerae, Flavobacterium ferrugineum et Mycobacterium tuberculosis. Ces Mabs pourraient être utilisés pour la détection de E. tarda chez les poisons.

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