title: Evaluation of cocktails with recombinant proteins of Mycobacterium bovis for a specific diagnosis of bovine tuberculosis.
abstract: The Delayed type hypersensitivity skin test (DTH) and interferon-gamma assay are used for the diagnosis of bovine tuberculosis (TBB). The specificity of these diagnoses, however, is compromised because both are based on the response against purified protein derivative of Mycobacterium bovis (PPD-B). In this study, we assessed the potential of two cocktails containing M. bovis recombinant proteins: cocktail 1 (C1): ESAT-6, CFP-10 and MPB83 and cocktail 2 (C2): ESAT-6, CFP-10, MPB83, HspX, TB10.3, and MPB70. C1, C2, and PPD-B showed similar response by DTH in M. bovis-sensitized guinea pigs. Importantly, C1 induced a lower response than PPD-B in M. avium-sensitized guinea pigs. In cattle, C1 displayed better performance than PPD-B and C2; indeed, C1 showed the least detection of animals either vaccinated or Map-infected. To optimize the composition of the cocktails, we obtained protein fractions from PPD-B and tested their immunogenicity in experimentally M. bovis-infected cattle. In one highly reactive fraction, seven proteins were identified. The inclusion of FixB in C1 enhanced the recognition of M. bovis-infected cattle without compromising specificity. Our data provide a promising basis for the future development of a cocktail for TBB detection without interference by the presence of sensitized or infected animals with other mycobacteria.
authors.affiliation: Instituto de Biotecnología, INTA, Los Reseros y Nicolás Repetto, 1686 Hurlingham, Buenos Aires, Argentina.
authors.forename: María Laura
authors.initials: ML
authors.lastname: Mon
authors.affiliation: Instituto de Biotecnología, INTA, Los Reseros y Nicolás Repetto, 1686 Hurlingham, Buenos Aires, Argentina.
authors.forename: Roberto Damián
authors.initials: RD
authors.lastname: Moyano
authors.affiliation: Instituto de Biotecnología, INTA, Los Reseros y Nicolás Repetto, 1686 Hurlingham, Buenos Aires, Argentina.
authors.forename: Mariana Noelia
authors.initials: MN
authors.lastname: Viale
authors.affiliation: Instituto de Biotecnología, INTA, Los Reseros y Nicolás Repetto, 1686 Hurlingham, Buenos Aires, Argentina.
authors.forename: María Alejandra
authors.initials: MA
authors.lastname: Colombatti Olivieri
authors.affiliation: INTA, Estación Experimental Agropecuaria Delta, 1670 Delta, Buenos Aires, Argentina.
authors.forename: Ignacio José
authors.initials: IJ
authors.lastname: Gamietea
authors.affiliation: Instituto de Biotecnología, INTA, Los Reseros y Nicolás Repetto, 1686 Hurlingham, Buenos Aires, Argentina.
authors.forename: Valeria Noely
authors.initials: VN
authors.lastname: Montenegro
authors.affiliation: SENASA, Dirección de Laboratorio y Control Técnico (DILAB), 1640 Martínez, Buenos Aires, Argentina.
authors.forename: Bernardo
authors.initials: B
authors.lastname: Alonso
authors.affiliation: Instituto de Biotecnología, INTA, Los Reseros y Nicolás Repetto, 1686 Hurlingham, Buenos Aires, Argentina.
authors.forename: María de la Paz
authors.initials: Mde L
authors.lastname: Santangelo
authors.affiliation: LIONEX Diagnostics & Therapeutics GmbH, 38126 Braunschweig, Germany.
authors.forename: Mahavir
authors.initials: M
authors.lastname: Singh
authors.affiliation: Unidad de Bioquímica y Proteómica Analítica, Institut Pasteur de Montevideo/Instituto de Investigaciones Biológicas Clemente Estable, 11400 Montevideo, Uruguay.
authors.forename: Rosario
authors.initials: R
authors.lastname: Duran
authors.affiliation: Instituto de Biotecnología, INTA, Los Reseros y Nicolás Repetto, 1686 Hurlingham, Buenos Aires, Argentina.
authors.forename: María Isabel
authors.initials: MI
authors.lastname: Romano
mesh_terms: D000818:Animals; D001500:BCG Vaccine / Q000276:immunology; D002417:Cattle; D006168:Guinea Pigs; D006968:Hypersensitivity, Delayed / Q000276:immunology / Q000382:microbiology; D007371:Interferon-gamma / Q000378:metabolism; D013058:Mass Spectrometry; D009163:Mycobacterium bovis / Q000502:physiology*; D011994:Recombinant Proteins / Q000276:immunology; D014380:Tuberculosis, Bovine / Q000175:diagnosis* / Q000276:immunology / Q000382:microbiology*; D014611:Vaccination
chemical_list: D001500:BCG Vaccine; D011994:Recombinant Proteins; D007371:Interferon-gamma
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