title: Use of ELISA based on NcSRS2 of Neospora caninum expressed in Pichia pastoris for diagnosing neosporosis in sheep and dogs.
abstract: Neosporosis is a disease caused by the protozoon Neospora caninum that leads to significant economic losses in many countries. In the present study, we report on use of the recombinant protein NcSRS2 of N. caninum expressed in Pichia pastoris in an indirect immunoenzymatic assay (ELISA) for diagnosing neosporosis infection in sheep and dogs. We observed that the ELISA test yielded specificity of 94.5% and sensitivity of 100% for sheep and specificity of 93.3% and sensitivity of 100% for dogs. We observed that the sensitivity was higher than shown by the indirect fluorescent antibody test, and this was confirmed by means of Western blot. The results from this study suggest that the recombinant protein expressed in P. pastoris is a suitable antigen for use in immunodiagnosis to detect N. caninum in two important species exposed to this parasitosis.
authors.affiliation: Departamento de Microbiologia e Parasitologia, Instituto de Biologia, Universidade Federal de Pelotas, Pelotas, RS, Brazil.
authors.forename: Amanda Fernandes
authors.initials: AF
authors.lastname: Pinheiro
authors.affiliation: Núcleo de Tecnologia, Centro de Desenvolvimento Tecnológico, Universidade Federal de Pelotas, Pelotas, RS, Brazil.
authors.forename: Sibele
authors.initials: S
authors.lastname: Borsuk
authors.affiliation: Departamento de Microbiologia e Parasitologia, Instituto de Biologia, Universidade Federal de Pelotas, Pelotas, RS, Brazil.
authors.forename: Maria Elisabeth Aires
authors.initials: ME
authors.lastname: Berne
authors.affiliation: Núcleo de Tecnologia, Centro de Desenvolvimento Tecnológico, Universidade Federal de Pelotas, Pelotas, RS, Brazil.
authors.forename: Luciano da Silva
authors.initials: Lda S
authors.lastname: Pinto
authors.affiliation: Centro Nacional Gado de Corte, Empresa Brasileira de Pesquisa Agropecuária, Campo Grande, MS, Brazil.
authors.forename: Renato
authors.initials: R
authors.lastname: Andreotti
authors.affiliation: Departamento de Microbiologia e Parasitologia, Instituto de Biologia, Universidade Federal de Pelotas, Pelotas, RS, Brazil.
authors.forename: Talita
authors.initials: T
authors.lastname: Roos
authors.affiliation: Núcleo de Tecnologia, Centro de Desenvolvimento Tecnológico, Universidade Federal de Pelotas, Pelotas, RS, Brazil.
authors.forename: Barbara Couto
authors.initials: BC
authors.lastname: Roloff
authors.affiliation: Departamento de Microbiologia e Parasitologia, Instituto de Biologia, Universidade Federal de Pelotas, Pelotas, RS, Brazil.
authors.forename: Fábio Pereira Leivas
authors.initials: FP
authors.lastname: Leite
mesh_terms: D000818:Animals; D000953:Antigens, Protozoan / Q000096:biosynthesis / Q000097:blood* / Q000276:immunology; D000954:Antigens, Surface / Q000096:biosynthesis / Q000097:blood* / Q000276:immunology; D004283:Dog Diseases / Q000097:blood* / Q000175:diagnosis / Q000469:parasitology*; D004285:Dogs; D004797:Enzyme-Linked Immunosorbent Assay; D018688:Neospora / Q000276:immunology; D010843:Pichia / Q000378:metabolism; D011529:Protozoan Infections, Animal / Q000097:blood* / Q000175:diagnosis; D015800:Protozoan Proteins / Q000096:biosynthesis / Q000097:blood* / Q000276:immunology; D012756:Sheep; D012757:Sheep Diseases / Q000097:blood* / Q000175:diagnosis / Q000469:parasitology*
chemical_list: D000953:Antigens, Protozoan; D000954:Antigens, Surface; D015800:Protozoan Proteins; C115389:SRS2 protein, Neospora caninum