ELISA detection of bovine viral diarrhoea virus specific antibodies using recombinant antigen and monoclonal antibodies |
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| Authors: | C Lecomte J J Pin L De Moerlooze D Vandenbergh A F Lambert P P Pastoret G Chappuis |
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| Institution: | Eurogentec, Campus du Sart Tilman, Liège, Belgium. |
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| Abstract: | A panel of monoclonal antibodies was prepared by immunization of BALB/c mice with Moredun (BD) virus strains. These antibodies were characterized by immunofluorescence and seroneutralization against BD, BVD and hog cholera (HC) virus strains, and radioimmunoprecipitation of BVD-infected cells extracts. The MAbs reacting with the majority of the Pestivirus strains recognize the 80 kDa antigen of the BVD cytophathic strains. The 80 kDa antigen of the BVD/Osloss virus strain has been cloned and expressed in E. coli as a fusion protein with beta-galactosidase. The fusion protein has been purified from inclusion bodies and used successfully as an antigen for ELISA detection of BVDV specific antibodies in bovine sera. A competitive ELISA using MAbs is more specific than a direct assay. These results compare well with the ones obtained with antigen extracted from BVDV-infected cells. |
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