Desarrollo de pruebas serológicas para la identificación de ovinos inmunizados con vacunas-GFP

• A. Zablaza ^[1] ; B. San Román ^[1] ; P.M. Muñoz ^[1] ; J.M. Blasco ^[1] ; M.J. Grilló ^[1]
  1. [1] Gobierno de Navarra

     Gobierno de Navarra

     Pamplona, España

     
• Localización: XV Jornadas sobre Producción Animal: 14 y 15 de mayo de 2013, Zaragoza / Jorge Hugo Calvo Lacosta (aut.), Isabel Casasús Pueyo (aut.), Margalida Joy Torrens (aut.), Javier Álvarez Rodríguez (aut.), Luis Varona Aguado (aut.), Begoña Panea Doblado (aut.), Carlos Calvete Margolles (aut.), Joaquim Barcells Teres (aut.), Vol. 2, 2013, ISBN 978-84-695-7684-7, págs. 831-833
• Idioma: español
• Títulos paralelos:
  • Development of serological tests for identifying sheep immunized with GFP-vaccines
• Enlaces
  • Texto Completo Libro  1 (pdf)   2 (pdf) 
• Resumen

  • Serological interference in diagnostic tests is a major problem associated to the use of veterinary vaccines, hampering the differentiation between infected and vaccinated animals (DIVA). To solve this in the ovine brucellosis context, our group is developing new Brucella vaccines tagged with green fluorescent protein (GFP) as a xenogenic marker, to be used in combination with both classical and GFP DIVA-tests. The aim of this work was to develop serological tests (DDG-GFP and ELISA-GFP) to detect GFP antibodies in sheep. The optimal concentration of the proteins in DDG-GFP was that containing 31.25 μg/mL of GFP. In ELISA-GFP, GFP-GST fusion protein was more suitable than GFP as antigen. Moreover, other ELISA-GFP factors (blocking effect, serum and conjugate optimal dilutions, and time of ABTS substrate incubation) were evaluated, setting up an ELISA-GFP detecting all hyperimmune sera, and showing 99 % specificity when 92 negative sheep sera were tested. In conclusion, the serologic tests developed allowed to detect anti-GFP sheep antibodies, and could be useful along with recombinant GFP vaccines, as a DIVA strategy.