Ana Paula Gori Palka1,2,3, Tatiana Reichert Assunção de Matos1,4, Claudemir de Souza1,3, Danilo Santos Eugênio1,3, Marco Aurélio Krieger1,3,4, Stenio Perdigão Fragoso1,4, Daniela Parada Pavoni5,6,7. 1. Instituto Carlos Chagas/ICC, Fundação Oswaldo Cruz/Fiocruz, Curitiba, PR, Brazil. 2. Instituto de Tecnologia do Paraná/Tecpar, Curitiba, PR, Brazil. 3. Programa de Pós-graduação em Biologia Celular e Molecular, Universidade Federal do Paraná/UFPR, Curitiba, PR, Brazil. 4. Programa de Pós-graduação em Biociências e Biotecnologia, Instituto Carlos Chagas/ICC, Fundação Oswaldo Cruz/Fiocruz, Curitiba, PR, Brazil. 5. Instituto Carlos Chagas/ICC, Fundação Oswaldo Cruz/Fiocruz, Curitiba, PR, Brazil. daniela.pavoni@fiocruz.br. 6. Programa de Pós-graduação em Biologia Celular e Molecular, Universidade Federal do Paraná/UFPR, Curitiba, PR, Brazil. daniela.pavoni@fiocruz.br. 7. Programa de Pós-graduação em Biociências e Biotecnologia, Instituto Carlos Chagas/ICC, Fundação Oswaldo Cruz/Fiocruz, Curitiba, PR, Brazil. daniela.pavoni@fiocruz.br.
Abstract
BACKGROUND: Infectious bursal disease (IBD), also known as Gumboro disease, is a viral infection that causes mortality and immunosuppression in chickens (Gallus gallus). VP2 and VP3 are the major structural viral capsid components and are the most immunogenic proteins of IBD virus (IBDV). Reliable diagnostic tests using VP2 and VP3 produced in heterologous systems are important tools to control this infection. One advantage of an IBD diagnostic based on VP3, over those that use VP2, is that VP3 has linear epitopes, enabling its production in bacteria. RESULTS: We tested the suitability of recombinant VP3 (rVP3) as a diagnostic reagent in an enzyme-linked immunosorbent assay (ELISA). Compared with a commercial test, rVP3 ELISA showed high sensitivity and specificity as a diagnostic tool for vaccinated animals. In addition, rVP3, but not the commercial ELISA, was able to detect antibodies in nonvaccinated chickens, probably developed against circulating IBDV strains. It was possible the assessment of VP3 regions antigenicity using chicken antisera. CONCLUSIONS: The full-length recombinant VP3 can be used to assess post vaccination immunological status of chickens and its production is feasible and inexpensive. The evaluation of VP3 regions as candidates for general use in the diagnosis of IBD in chickens should be conducted with caution. Our work was the first to identify several regions of VP3 recognized by chicken antibodies.
BACKGROUND:Infectious bursal disease (IBD), also known as Gumboro disease, is a viral infection that causes mortality and immunosuppression in chickens (Gallus gallus). VP2 and VP3 are the major structural viral capsid components and are the most immunogenic proteins of IBD virus (IBDV). Reliable diagnostic tests using VP2 and VP3 produced in heterologous systems are important tools to control this infection. One advantage of an IBD diagnostic based on VP3, over those that use VP2, is that VP3 has linear epitopes, enabling its production in bacteria. RESULTS: We tested the suitability of recombinant VP3 (rVP3) as a diagnostic reagent in an enzyme-linked immunosorbent assay (ELISA). Compared with a commercial test, rVP3 ELISA showed high sensitivity and specificity as a diagnostic tool for vaccinated animals. In addition, rVP3, but not the commercial ELISA, was able to detect antibodies in nonvaccinated chickens, probably developed against circulating IBDV strains. It was possible the assessment of VP3 regions antigenicity using chicken antisera. CONCLUSIONS: The full-length recombinant VP3 can be used to assess post vaccination immunological status of chickens and its production is feasible and inexpensive. The evaluation of VP3 regions as candidates for general use in the diagnosis of IBD in chickens should be conducted with caution. Our work was the first to identify several regions of VP3 recognized by chicken antibodies.