CLONING, SEQUENCING, EXPRESSION AND ANTIGENIC CHARACTERIZATION OF NCROP2 AND EVALUATION OF AN ELISA.
Autor(es): Maria Paula de Carvalho Ewald, Mario Augusto Ono, Carolina Nachi Rossi, João Luis Garcia, Marilda Carlos Vidotto
CLONING, SEQUENCING, EXPRESSION AND ANTIGENIC CHARACTERIZATION OF NCROP2 AND EVALUATION OF AN ELISA.
» Área de pesquisa: PROTOZOOLOGIA
» Instituição: Universidade Estadual de Londrina
» Agência de fomento e patrocinadores: Capes
Neospora caninum is a protozoa parasite that causes neosporosis, which is one of the most important abortifacient in cattle worldwide. The present work describes the isolation and purification of rROP2 from N. caninum, and their use in the development of an immunoassay for the diagnostic of anti-N. caninum antibodies from natural infected cattle. After expression of the rNcROP2 and cell disruption, it was treated with urea and purified by immobilized metal affinity chromatography. An indirect enzyme-linked immunosorbent assay (ELISA) was developed using rROP2 purified and the ELISA results were compared with indirect fluorescence antibody test (IFAT). In our search, the ELISA-ROP2 format did not discriminated positive and negative serum, indicating that rROP2 is not a good antigen for serological assay for diagnostic of Neosporosis. The Wester Blotting results showed that both positive and negative antisera IFAT-Nc do not react with the purified protein. The reactivity in both cases was very low.
