Development and application of an enzyme-linked immunosorbent assay for detection of bovine viral diarrhea antibody based on Erns glycoprotein expressed in a baculovirus system.
The BVDV envelope glycoprotein E(rns)/gp48 and the C terminal 79 amino acids of the capsid protein coding region were expressed in a baculovirus system and antigenically characterized. Western blot assay was used to detect recombinant E(rns) (r-E(rns)) in infected insect cells using specific monoclonal antibodies. The r-E(rns) was then used in an indirect ELISA to detect BVDV specific antibodies in a panel of 540 well-characterized sera. Results of the r-E(rns) ELISA were compared to those obtained with a commercially available competitive ELISA targeting anti-NS2/3 antibodies. A good correlation was observed between the 2 ELISA (kappa = 0.916, 95% C.I.: 0.876, 0.956). Using the commercial NS2/3 ELISA as the reference test, the relative sensitivity of r-E(rns) ELISA was 97.5% (95% C.I.: 94.3%, 99.1%) and the relative specificity was 93.9% (95% C.I.: 89.4%, 96.9%), while relative specificity was 100% (95% C.I.: 97%, 100%) using true negative sera (derived from a negative herd). All but 1 antigen positive animals (n = 36) tested negative in the r-E(rns) ELISA; among them all 22 confirmed PI animals were negative by r-E(rns) ELISA. The ability of r-E(rns) ELISA to identify cattle immunized with inactivated vaccine was also demonstrated in a small group of cattle, compared to an NS2/3 antibody ELISA. Results suggest that r-E(rns) ELISA represents an alternative test for antibody generated by natural infection or BVDV vaccination.
Grego
E
E
Dipartimento di Produzioni Animali, Epidemiolgia ed Ecologia, Facoltà di Medicina Veterinaria, Via Leonardo da Vinci 44, 10095 Grugliasco (TO), Italy.
Uslenghi
F
F
Strasser
M
M
Luzzago
C
C
Frigerio
M
M
Peletto
S
S
Rosati
S
S
D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology; D000914:Antibodies, Viral / Q000032:analysis* / Q000276:immunology*; D000956:Antigens, Viral / Q000276:immunology; D016367:Baculoviridae; D001912:Bovine Virus Diarrhea-Mucosal Disease / Q000175:diagnosis / Q000821:virology; D002417:Cattle; D002460:Cell Line; D001908:Diarrhea Viruses, Bovine Viral / Q000276:immunology*; D004797:Enzyme-Linked Immunosorbent Assay / Q000662:veterinary*; D006023:Glycoproteins / Q000235:genetics* / Q000276:immunology; D009036:Moths / Q000166:cytology; D011994:Recombinant Proteins; D013997:Time Factors; D015678:Viral Structural Proteins / Q000276:immunology
D000911:Antibodies, Monoclonal; D000914:Antibodies, Viral; D000956:Antigens, Viral; D006023:Glycoproteins; D011994:Recombinant Proteins; D015678:Viral Structural Proteins