Development and evaluation of an enzyme-linked immunosorbent assay for the serological diagnosis of the bovine herpesvirus 1 infection

Authors

  • Melissa C. Ferreira Universidade Estadual de Londrina
  • Kerlei C. Médici Universidade Estadual de Londrina
  • Alice F. Alfieri Universidade Estadual de Londrina
  • Amauri A. Alfieri Universidade Estadual de Londrina

DOI:

https://doi.org/10.5433/1679-0359.2005v26n3p363

Keywords:

Cattle, Bovine herpesvirus 1, Serology, Indirect ELISA.

Abstract

An indirect enzyme-linked immunosorbent assay (ELISA) using non-purified antigen was developed for serological diagnosis of the bovine herpesvirus 1 (BoHV-1) infection. Some blocking substances to prevent nonspecific reactions were evaluated in different concentrations and combinations. The best results were obtained using 5% skim milk powder as blocking solution and the serum/conjugated buffer added with 10% MDBK cells, 10% equine serum, 1% skim milk powder. The system was evaluated with a collection of positives (n=60) and negatives (n=62) bovine serum previously analyzed by seroneutralization (SN) technique for the BoHV-1 antibodies. The indirect ELISA standardized showed 98.3% of sensitivity and 95.2% of specificity (kappa: 0.93) when compared with the SN technique. These results would allow its implantation in the laboratorial routine for the serological diagnosis of the BoHV-1 infection. The use of non-purified antigen in ELISA facilitate the elaboration, reduces the production cost and makes possible the application of these technique in seroepidemiological study of the BoHV-1 infection in cattle herds.

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Author Biographies

Melissa C. Ferreira, Universidade Estadual de Londrina

Aluna do Programa de Pós-graduação em Ciência Animal (Área de Concentração: Sanidade Animal; Nível: Mestrado), Centro de Ciências Agrárias (CCA), Universidade Estadual de Londrina (UEL).

Kerlei C. Médici, Universidade Estadual de Londrina

Laboratório de Virologia Animal, Departamento de Medicina Veterinária Preventiva, CCA, UEL, Campus Universitário, caixa postal 6001, CEP. 86051-990, Londrina, Paraná, Brasil.

Alice F. Alfieri, Universidade Estadual de Londrina

Laboratório de Virologia Animal, Departamento de Medicina Veterinária Preventiva, CCA, UEL, Campus Universitário, caixa postal 6001, CEP. 86051-990, Londrina, Paraná, Brasil.

Amauri A. Alfieri, Universidade Estadual de Londrina

Laboratório de Virologia Animal, Departamento de Medicina Veterinária Preventiva, CCA, UEL, Campus Universitário, caixa postal 6001, CEP. 86051-990, Londrina, Paraná, Brasil.

Published

2005-06-30

How to Cite

Ferreira, M. C., Médici, K. C., Alfieri, A. F., & Alfieri, A. A. (2005). Development and evaluation of an enzyme-linked immunosorbent assay for the serological diagnosis of the bovine herpesvirus 1 infection. Semina: Ciências Agrárias, 26(3), 363–372. https://doi.org/10.5433/1679-0359.2005v26n3p363

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