title: In silico identification of Corynebacterium pseudotuberculosis antigenic targets and application in immunodiagnosis.
abstract: Caseous lymphadenitis (CLA) is a disease caused by Corynebacterium pseudotuberculosis. It affects mainly small ruminants and causes significant economic losses worldwide. Because symptoms are not immediately noticeable, CLA clinical diagnosis is not effective. Numerous serological tests are being developed to detect the disease in asymptomatic animals, but currently available immunoassays have problems with sensitivity. Current ELISA formats use native bacterial antigens, and recombinant proteins could be useful for improving the immunoassay parameters. The C. pseudotuberculosis proteins CP0126a, CP0369 and CP1957 were identified from 2097 candidate proteins by mature epitope density (MED) analysis, expressed in Escherichia coli and evaluated in an indirect immunoenzymic system. The CP0126a, CP0369 and CP1957 ELISAs showed 77.5 %, 92.5 % and 92.5 % specificity and 95 %, 90 % and 85 % sensitivity, respectively. Receiver operating characteristic (ROC) curve analysis showed an area under the curve of 0.874, 0.951 and 0.881, respectively. The proteins identified in silico were recognized by antibodies in the sera from infected animals without being recognized in negative samples. The ELISA assay using the rCP0369 protein as antigen had the greatest specificity and sensitivity values, followed by rCP1957. This is an interesting strategy for seroepidemiological investigations in sheep flocks due to its significant specificity and sensitivity.
authors.affiliation: Laboratório de Pesquisa em Doenças Infecciosas, Centro de Desenvolvimento Tecnológico, Biotecnologia, UFPel, Pelotas, RS 96010-900, Brazil.
authors.forename: Andrea de Fátima Silva
authors.initials: AFS
authors.lastname: Rezende
authors.affiliation: Laboratório de Pesquisa em Doenças Infecciosas, Centro de Desenvolvimento Tecnológico, Biotecnologia, UFPel, Pelotas, RS 96010-900, Brazil.
authors.forename: Alexandre Antunes
authors.initials: AA
authors.lastname: Brum
authors.affiliation: Laboratório de Pesquisa em Doenças Infecciosas, Centro de Desenvolvimento Tecnológico, Biotecnologia, UFPel, Pelotas, RS 96010-900, Brazil.
authors.forename: Carlos Guilherme
authors.initials: CG
authors.lastname: Reis
authors.affiliation: Laboratório de Pesquisa em Doenças Infecciosas, Centro de Desenvolvimento Tecnológico, Biotecnologia, UFPel, Pelotas, RS 96010-900, Brazil.
authors.forename: Henrique Ramos
authors.initials: HR
authors.lastname: Angelo
authors.affiliation: Laboratório de Pesquisa em Doenças Infecciosas, Centro de Desenvolvimento Tecnológico, Biotecnologia, UFPel, Pelotas, RS 96010-900, Brazil.
authors.forename: Karen Silva
authors.initials: KS
authors.lastname: Leal
authors.affiliation: Laboratório de Pesquisa em Doenças Infecciosas, Centro de Desenvolvimento Tecnológico, Biotecnologia, UFPel, Pelotas, RS 96010-900, Brazil.
authors.forename: Mara Thais de Oliveira
authors.initials: MTO
authors.lastname: Silva
authors.affiliation: Laboratório de Biologia Molecular, Faculdade de Ciências Ambientais, UFGD, Dourados, MS 79825-070, Brazil.
authors.forename: Simone
authors.initials: S
authors.lastname: Simionatto
authors.affiliation: Laboratório de Genética Celular e Molecular, Instituto de Ciências Biológicas, UFMG, Belo Horizonte, MG 31270-901, Brazil.
authors.forename: Vasco
authors.initials: V
authors.lastname: Azevedo
authors.affiliation: Laboratório de Genética Celular e Molecular, Instituto de Ciências Biológicas, UFMG, Belo Horizonte, MG 31270-901, Brazil.
authors.forename: Anderson
authors.initials: A
authors.lastname: Santos
authors.affiliation: Laboratório de Imunologia e Biologia Molecular, Instituto de Ciências da Saúde, UFBA, Salvador, BA 40140-100, Brazil.
authors.forename: Ricardo Wagner
authors.initials: RW
authors.lastname: Portela
authors.affiliation: Laboratório de Pesquisa em Doenças Infecciosas, Centro de Desenvolvimento Tecnológico, Biotecnologia, UFPel, Pelotas, RS 96010-900, Brazil.
authors.forename: Odir
authors.initials: O
authors.lastname: Dellagostin
authors.affiliation: Laboratório de Pesquisa em Doenças Infecciosas, Centro de Desenvolvimento Tecnológico, Biotecnologia, UFPel, Pelotas, RS 96010-900, Brazil.
authors.forename: Sibele
authors.initials: S
authors.lastname: Borsuk
mesh_terms: D000942:Antigens, Bacterial / Q000737:chemistry* / Q000378:metabolism; D001426:Bacterial Proteins / Q000737:chemistry / Q000378:metabolism; D003198:Computer Simulation; D003354:Corynebacterium Infections / Q000175:diagnosis* / Q000382:microbiology*; D016925:Corynebacterium pseudotuberculosis; D004797:Enzyme-Linked Immunosorbent Assay; D018604:Epitope Mapping; D006801:Humans; D007159:Immunologic Tests / Q000379:methods*; D012680:Sensitivity and Specificity
chemical_list: D000942:Antigens, Bacterial; D001426:Bacterial Proteins