BVD CONSULT

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

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Abstract

Correspondence: Corresponding Author: Sergio Rosati, Dipartimento di Produzioni Animali, Epidemiolgia ed Ecologia, Facoltà di Medicina Veterinaria, Via Leonardo da Vinci 44, 10095 Grugliasco (TO), Italy, e-mail: sergio.rosati@unito.it

The BVDV envelope glycoprotein Erns/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 Erns (r-Erns) in infected insect cells using specific monoclonal antibodies. The r-Erns was then used in an indirect ELISA to detect BVDV specific antibodies in a panel of 540 well-characterized sera. Results of the r-Erns 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-Erns 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-Erns ELISA; among them all 22 confirmed PI animals were negative by r-Erns ELISA. The ability of r-Erns 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-Erns ELISA represents an alternative test for antibody generated by natural infection or BVDV vaccination.

Key Words: Baculovirus • bovine viral diarrhea virus • ELISA • Erns glycoprotein

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Grego E, Uslenghi F, Strasser M, Luzzago C, et al.: J Vet Diagn Invest 19: 21-27, 2007.