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Journal of Clinical Microbiology, August 2003, p. 3675-3680, Vol. 41, No. 8
0095-1137/03/$08.00+0     DOI: 10.1128/JCM.41.8.3675-3680.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.

Detection of Circulating gp43 Antigen in Serum, Cerebrospinal Fluid, and Bronchoalveolar Lavage Fluid of Patients with Paracoccidioidomycosis

Silvia Helena Marques da Silva,1 Arnaldo Lopes Colombo,2 Maria Heloisa Souza Lima Blotta,3 José Daniel Lopes,1 Flávio Queiroz-Telles,4 and Zoilo Pires de Camargo1*

Department of Microbiology, Immunology and Parasitology,1 Department of Infectious Diseases, Federal University of São Paulo, São Paulo,2 Department of Clinical Pathology, Medical School, State University of Campinas, Campinas, São Paulo,3 Department of Community Health, Medical School, Federal University of Paraná, Curitiba, Paraná, Brazil4

Received 17 January 2003/ Returned for modification 14 April 2003/ Accepted 16 May 2003

Paracoccidioidomycosis (PCM) is an important systemic fungal disease, particularly among individuals living and working in rural areas of endemicity in Latin America, who, without antifungal therapy, may develop fatal acute or chronic infection. For such patients, the detection of antibody responses by immunodiffusion is of limited value due to false-negative results. In contrast, the detection of Paracoccidioides brasiliensis gp43 circulating antigen may represent a more practical approach to the rapid diagnosis of the disease. Accordingly, an inhibition enzyme-linked immunosorbent assay (inh-ELISA) was developed for the detection of a 43-kDa P. brasiliensis-specific epitope incorporating a species-specific murine monoclonal antibody. With sera from patients with acute and chronic forms of the disease (n = 81), the overall sensitivity of the test was found to be 95.1%, while specificity was found to be 97.5% compared to that with normal human sera from blood donors (n = 93) and sera from patients with other chronic fungal infections (histoplasmosis [n = 33] and cryptococcosis [n = 20]). The inh-ELISA detected circulating antigen in 100% of patients with the acute form of PCM and in 95.31 and 100% of patients with the chronic multifocal and unifocal forms of PCM according to the patient's clinical presentation. Cerebrospinal fluid from 14 patients with neuroparacoccidioidomycosis and 13 samples of bronchoalveolar lavage fluid from patients with pulmonary unifocal PCM were also tested for gp43 detection, with the test showing 100% sensitivity and specificity. This novel, highly specific inh-ELISA represents a significant addition to the existing tests for the diagnosis of PCM.

* Corresponding author. Mailing address: Universidade Federal de São Paulo, Disciplina de Biologia Celular, 04023-062, Rua Botucatu 862/8° andar, São Paulo, SP, Brazil. Phone: 55 11 5576 4523. Fax: 55 11 5571 58 77. E-mail: zoilo{at}ecb.epm.br.

Journal of Clinical Microbiology, August 2003, p. 3675-3680, Vol. 41, No. 8
0095-1137/03/$08.00+0     DOI: 10.1128/JCM.41.8.3675-3680.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.



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