ELISA detection of bovine viral diarrhoea virus specific antibodies using recombinant antigen and monoclonal antibodies.
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.
Lecomte
C
C
Eurogentec, Campus du Sart Tilman, Liège, Belgium.
Pin
J J
JJ
De Moerlooze
L
L
Vandenbergh
D
D
Lambert
A F
AF
Pastoret
P P
PP
Chappuis
G
G
D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology*; D000914:Antibodies, Viral / Q000032:analysis*; D000918:Antibody Specificity; D000956:Antigens, Viral / Q000276:immunology*; D001667:Binding, Competitive; D002417:Cattle; D001908:Diarrhea Viruses, Bovine Viral / Q000276:immunology*; D004797:Enzyme-Linked Immunosorbent Assay; D005455:Fluorescent Antibody Technique; D006825:Hybridomas; D051379:Mice; D008807:Mice, Inbred BALB C; D009500:Neutralization Tests; D010576:Pestivirus / Q000276:immunology*; D015531:Radioimmunoprecipitation Assay; D011994:Recombinant Proteins / Q000276:immunology
D000911:Antibodies, Monoclonal; D000914:Antibodies, Viral; D000956:Antigens, Viral; D011994:Recombinant Proteins