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Clinical and Diagnostic Laboratory Immunology, March 2002, p. 374-377, Vol. 9, No. 2
1071-412X/02/$04.00+0     DOI: 10.1128/CDLI.9.2.374-377.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.

Heterologous Expression, Purification, and Immunological Reactivity of a Recombinant HSP60 from Paracoccidioides brasiliensis

Daniela A. Cunha,1 Roseli M. Zancopé-Oliveira,2 M. Sueli, S. Felipe,3 Silvia M. Salem-Izacc,1 George S. Deepe Jr.,4 and Célia M. A. Soares1*

Laboratório de Biologia Molecular, ICB, Universidade Federal de Goiás, 74001-970 Goiânia, Goiás,1 Laboratório de Micologia Médica, Hospital Evandro Chagas, Fundação Oswaldo Cruz, Rio de Janeiro,2 Laboratório de Biologia Molecular, Instituto de Biologia, Universidade de Brasília, Brasília D.F., Brazil,3 Division of Infectious Diseases, College of Medicine, University of Cincinnati, Cincinnati, Ohio 45267-05604

Received 26 July 2001/ Returned for modification 25 September 2001/ Accepted 20 November 2001

The complete coding cDNA of HSP60 from Paracoccidioides brasiliensis was overexpressed in an Escherichia coli host to produce high levels of recombinant protein. The protein was purified by affinity chromatography. A total of 169 human serum samples were tested for reactivity by Western blot analysis with the purified HSP60 recombinant protein. Immunoblots indicated that the recombinant P. brasiliensis HSP60 was recognized by antibodies in 72 of 75 sera from paracoccidioidomycosis patients. No cross-reactivity was detected with individual sera from patients with aspergillosis, sporotrichosis, cryptococcosis, and tuberculosis. Reactivity to HSP60 was observed in sera from 9.52% of control healthy individuals and 11.5% of patients with histoplasmosis. The high sensitivity and specificity (97.3 and 92.5%, respectively) for HSP60 suggested that the recombinant protein can be used singly or in association with other recombinant antigens to detect antibody responses in P. brasiliensis-infected patients.

* Corresponding author. Mailing address: Laboratório de Biologia Molecular, Instituto de Ciências Biológicas, ICBII, Campus II, Universidade Federal de Goiás, 74001-970, Goiânia, Goiás, Brazil. Phone: 55-62-5211110. Fax: 55-62-5211190. E-mail: celia{at}icb1.ufg.br.

Clinical and Diagnostic Laboratory Immunology, March 2002, p. 374-377, Vol. 9, No. 2
1071-412X/02/$04.00+0     DOI: 10.1128/CDLI.9.2.374-377.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.



This article has been cited by other articles:

  • Diez, S., Gomez, B. L., McEwen, J. G., Restrepo, A., Hay, R. J., Hamilton, A. J. (2003). Combined Use of Paracoccidioides brasiliensis Recombinant 27-Kilodalton and Purified 87-Kilodalton Antigens in an Enzyme-Linked Immunosorbent Assay for Serodiagnosis of Paracoccidioidomycosis. J. Clin. Microbiol. 41: 1536-1542 [Abstract] [Full Text]  


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