An ELISA was established to measure bovine IgG directed against the recombinant antigenic determinant of Nc-p43, a major surface antigen of Neospora caninum. In a previous study, two thirds of the C-terminal of the molecule was expressed as a 6 × His tagged protein (Ncp43P) for ELISA using 2/3 of the N-terminal of SAG1 from Toxoplasma gondii as a control (TgSAG1A). Among 852 cattle sera collected from stock farms scattered nation-wide, 103 sera (12.1%) were found to react with Ncp43P positively, but no positive reaction was observed with TgSAG1A. This study shows that Ncp43P could be available as an efficient antigen for the diagnosis of neosporosis in cattle. Furthermore, it together with TgSAG1A, could be useful for the differential diagnosis of N. caninum and T. gondii infections in other mammals.
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Use of ELISA based on NcSRS2 of Neospora caninumexpressed in Pichia pastoris for diagnosing neosporosis in sheep and dogs Amanda Fernandes Pinheiro, Sibele Borsuk, Maria Elisabeth Aires Berne, Luciano da Silva Pinto, Renato Andreotti, Talita Roos, Barbara Couto Roloff, Fábio Pereira Leivas Leite Revista Brasileira de Parasitologia Veterinária.2015; 24(2): 148. CrossRef
Diagnostic Potential of Anti-rNcp-43 Polyclonal Antibodies for the Detection of Neospora caninum Gizele Lima de Sá, Diene de Borba Pacheco, Leonardo Garcia Monte, Francine Alves Sinnott, Marina Amaral Xavier, Caroline Rizzi, Sibele Borsuk, Maria Elisabeth Aires Berne, Renato Andreotti, Cláudia Pinho Hartleben Current Microbiology.2014; 68(4): 472. CrossRef
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Epidemiology and Control of Neosporosis andNeospora caninum J. P. Dubey, G. Schares, L. M. Ortega-Mora Clinical Microbiology Reviews.2007; 20(2): 323. CrossRef
Diagnosis of bovine neosporosis: Recent advances and perspectives Luis Ortega-Mora, Aurora Fernández-García, Mercedes Gómez-Bautista Acta Parasitologica.2006; 51(1): 1. CrossRef
Diagnosis of bovine neosporosis J.P. Dubey, G. Schares Veterinary Parasitology.2006; 140(1-2): 1. CrossRef
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The antigenic domain of the major surface protein (Nc-p43) of Neospora caninum was examined by polymerase chain reaction of its gene fragments and recombinant expression as GST fusion proteins. The fragments of Nc-p43 were as follow: a total open reading frame (OFR), T; OFR without signal sequence and C-terminal hydrophobic sequence, S; N-terminal 2/3 parts of S, A; C-terminal 2/3 parts, P; N-terminal 1/3 part, X; middle 1/3 part, Y; and C-terminal 1/3 part, Z, respectively. The DNA fragments were cloned into pGEX-4T vector. Recombinant plasmids transformed into Escherichia coli of BL21 pLysS (DE3) strain were induced to express GST or GST fused fragments of Nc-p43 such as 69 kDa protein for T, 66 kDa for S, 52 kDa for A, 53 kDa for P, and 40 kDa proteins for X, Y, and Z, respectively in SDS-PAGE. The Nc-p43 fragments of T, S, and P reacted with a bovine serum of neosporosis while those of A, X, Y, and Z together with GST did not in the western blot. These findings suggest that the antigenic domain of Nc-p43 of N. caninum may be localized in the C-terminal 2/3 parts. Together with A19 clone in SAG1 of Toxoplasma gondii (Nam et al., 1996), the P fragment of Nc-p43 could be used as efficient antigens to diagnose and differentiate those infections with both species.
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