Production of monoclonal antibodies specific to major outer membrane protein of Edwardsiella tarda.
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 44kDa 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 1x10(4)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.
Kumar
Gokhlesh
G
National Bureau of Fish Genetic Resources, Lucknow, Uttar Pradesh, India.
Rathore
Gaurav
G
Sengupta
U
U
Kapoor
D
D
Lakra
W S
WS
D000818:Animals; D000907:Antibodies, Bacterial / Q000096:biosynthesis* / Q000276:immunology; D000911:Antibodies, Monoclonal / Q000096:biosynthesis* / Q000276:immunology; D001425:Bacterial Outer Membrane Proteins / Q000737:chemistry / Q000276:immunology* / Q000302:isolation & purification; D015153:Blotting, Western; D003429:Cross Reactions; D020609:Edwardsiella tarda / Q000276:immunology* / Q000302:isolation & purification; D004756:Enterobacteriaceae Infections / Q000175:diagnosis / Q000276:immunology / Q000382:microbiology / Q000662:veterinary*; D004797:Enzyme-Linked Immunosorbent Assay; D005393:Fish Diseases / Q000175:diagnosis* / Q000276:immunology / Q000382:microbiology; D006801:Humans; D006825:Hybridomas / Q000378:metabolism; D007074:Immunoglobulin G / Q000096:biosynthesis / Q000276:immunology; D007194:India; D051379:Mice; D010473:Perciformes; D012680:Sensitivity and Specificity
D000907:Antibodies, Bacterial; D000911:Antibodies, Monoclonal; D001425:Bacterial Outer Membrane Proteins; D007074:Immunoglobulin G