Production of recombinant EMA-1 protein and its application for the diagnosis of Theileria equi using an enzyme immunoassay in horses from Sao Paulo State, Brazil

Autor(es)

Baldani, Cristiane Divan ^[1] ; Hilario, Eduardo ^[2] ; Higa Nakaghi, Andrea Cristina ^[3] ; Bertolini, Maria Celia ^[4] ; Machado, Rosangela Zacarias ^[3]

Afiliação do(s) autor(es)

^[1] Univ Fed Rural Rio de Janeiro UFRRJ, Inst Vet, Dept Med & Cirurgia Vet, Rio De Janeiro - Brazil

^[2] Univ Calif Riverside, Dept Biochem, Riverside, CA 92521 - USA

^[3] Univ Estadual Paulista UNESP, Dept Patol Vet, Fac Ciencias Agr & Vet, BR-14884900 Jaboticabal, SP - Brazil

^[4] Univ Estadual Paulista UNESP, Dept Bioquim & Tecnol Quim, BR-14884900 Jaboticabal, SP - Brazil

• Tipo de documento: Artigo Científico
• Fonte: REVISTA BRASILEIRA DE PARASITOLOGIA VETERINARIA; v. 20, n. 1, p. 54-60, JAN-MAR 2011.
• Citações Web of Science: 4

Resumo

The erythrocytic-stage surface protein, Equi Merozoite Antigen 1 (EMA-1), is a major candidate for the development of a diagnostic antigen for equine piroplasmosis. In order to establish an effective diagnostic method for practical use, the gene encoding the entire EMA-1 of Theileria equi Jaboticabal strain was cloned and expressed in Escherichia coli as a histidine-tagged protein (His6-EMA1). The expressed EMA-1 reacted with specific antibodies in Western blot and had an apparent molecular mass of 34 kDa which was largely consistent with its theoretical value. The nucleotide sequence of the EMA-1 gene of Jaboticabal strain was comparatively analyzed with other published sequences. The results indicated a high degree of homology with EMA-1 genes of all other strains isolated from various countries. The recombinant purified His6-EMA1 protein was tested in an enzyme-linked immunosorbent assay (ELISA) for the detection of antibodies anti-T equi in horses. The ELISA clearly differentiated T. equi-infected from Babesia caballi-infected horse sera or normal horse sera. Field serum samples collected from horses in the State of Sao Paulo, Southeastern Brazil, were examined for the diagnosis of T. equi infection by ELISA. Of 170 samples analyzed, 95.88% (163/170) were positive for T. equi infection. These results suggest that the His6-EMA1 protein expressed in E. coli could be a reliable immunodiagnostic antigen for ELISA test and that T. equi infection is a serious concern in the State of Sao Paulo, Brazil. (AU)

• Processo FAPESP: 03/12793-3 - Avaliacao da reposta imune humoral em equinos naturalmente infectados por babesia equi pela utilizacao de antigeno recombinante ema-1 e antigenos parcialmente purificados e caracterizacao antigenica e...
• Beneficiário: Cristiane Divan Baldani
• Modalidade de apoio: Bolsas no Brasil - Doutorado
• Processo FAPESP: 04/05223-9 - Avaliação da resposta imune em equinos naturalmente infectados por Babesia equi pela utilização de antígeno recombinante EMA-1 e antígenos parcialmente purificados e caracterização antigênica e molecular da cepa, Jaboticabal - SP
• Beneficiário: Rosangela Zacarias Machado
• Modalidade de apoio: Auxílio à Pesquisa - Regular