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Biochemical Characterization of an In-House Coccidioides Antigen: Perspectives for the Immunodiagnosis of Coccidioidomycosis
Specialized Medical Mycology Center, School of Medicine, Federal University of Ceará, Fortaleza, CE, CEP, 60430-270, Brazil
Postgraduate Program in Veterinary Sciences, School of Veterinary Medicine, Ceará State University, Fortaleza, CE, CEP, 60740-000, Brazil
Department of Biochemistry and Molecular Biology, Federal University of Ceará, Pici Campus, Fortaleza, CE, 60455-760, Brazil
Department of Microbiology, Immunology and Parasitology, São Paulo Federal University, São Paulo, 04021-001, Brazil
* Author to whom correspondence should be addressed.
Received: 23 March 2012; in revised form: 18 June 2012 / Accepted: 20 June 2012 / Published: 28 June 2012
Abstract: The objective of this study was to evaluate the reactivity of an in-house antigen, extracted from a strain of C. posadasii isolated in northeastern Brazil, by radial immunodiffusion and Western blotting, as well as to establish its biochemical characterization. The protein antigen was initially extracted with the use of solid ammonium sulfate and characterized by 1-D electrophoresis. Subsequently, it was tested by means of double radial immunodiffusion and Western blotting. A positive reaction was observed against the antigen by both immunodiagnostic techniques tested on sera from patients suffering from coccidioidomycosis. Besides this, two immunoreactive protein bands were observed and were revealed to be a β-glucosidase and a glutamine synthetase after sequencing of the respective N-terminal regions. Our in-house Coccidioides antigen can be promising as a quick and low-cost diagnostic tool without the risk of direct manipulation of the microorganism.
Keywords: coccidioidomycosis; immunodiagnosis; antigens; β-glucosidase; glutamine
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Filho, R.E.M.; Bandeira, S.P.; Brillhante, R.S.N.; Rocha, M.F.G.; Vasconcelos, I.M.; Pereira, M.L.; Castelo-Branco, D.S.C.M.; Rocha, F.A.C.; Camargo, Z.P.; Ramos, M.V.; Cordeiro, R.A.; Sidrim, J.J.C. Biochemical Characterization of an In-House Coccidioides Antigen: Perspectives for the Immunodiagnosis of Coccidioidomycosis. Molecules 2012, 17, 7854-7863.
Filho REM, Bandeira SP, Brillhante RSN, Rocha MFG, Vasconcelos IM, Pereira ML, Castelo-Branco DSCM, Rocha FAC, Camargo ZP, Ramos MV, Cordeiro RA, Sidrim JJC. Biochemical Characterization of an In-House Coccidioides Antigen: Perspectives for the Immunodiagnosis of Coccidioidomycosis. Molecules. 2012; 17(7):7854-7863.
Filho, Renato Evando Moreira; Bandeira, Silviane Praciano; Brillhante, Raimunda Sâmia Nogueira; Rocha, Marcos Fábio Gadelha; Vasconcelos, Ilka Maria; Pereira, Mirella Leite; Castelo-Branco, Débora de Souza Collares Maia; Rocha, Francisco Airton Castro; Camargo, Zoilo Pires de; Ramos, Marcio Viana; Cordeiro, Rossana de Aguiar; Sidrim, José Júlio Costa. 2012. "Biochemical Characterization of an In-House Coccidioides Antigen: Perspectives for the Immunodiagnosis of Coccidioidomycosis." Molecules 17, no. 7: 7854-7863.