XI International Meeting on Paracoccidioidomycosis

XI International Meeting on Paracoccidioidomycosis

Resume:66-1

Poster (Painel)

66-1

Morphological aspects, protein and molecular profile of regional isolates of Paracoccidioides brasiliensis.

Authors:

Elaine Patrícia Tavares do Espírito Santo (UFPA/ICB/PPG-BAIP - Universidade Federal do Pará- UFPA/ICB/PPg-BAIPIEC/SVS/MS - Instituto Evandro Chagas- IEC/SVS/MS) ; Fabiola Silveira Gomes (UFPA/ICB/PPG-BAIP - Universidade Federal do Pará- UFPA/ICB/PPg-BAIPIEC/SVS/MS - Instituto Evandro Chagas- IEC/SVS/MS) ; Maurimélia Mesquita da Costa (UFPA/ICB/PPG-BAIP - Universidade Federal do Pará- UFPA/ICB/PPg-BAIPIEC/SVS/MS - Instituto Evandro Chagas- IEC/SVS/MS) ; Silvia Helena Marques da Silva (IEC/SVS/MS - Instituto Evandro Chagas- IEC/SVS/MSUFPA/ICB/PPG-BAIP - Universidade Federal do Pará- UFPA/ICB/PPg-BAIP)

Abstract

Paracoccidioides brasiliensis is the etiological agent of the paracoccidioidomycosis (PCM), one of the most important systemic mycosis in Central and South American. The diagnosis is usually made by isolating the fungus from the secretions of the lesions or serological tests for the detection of specific antibodies. The aim of this work was to describe the morphological features, specific antigens and molecular profile of regional isolates (Pará, north region of Brazil) of P. brasiliensis. We studied 2 clinical strains of P. brasiliensis (Pb WAL and Pb 1744) previously obtained from oral mucosal lesions of patients with chronic form, isolated in Pará, Brazil. Cellular extract from 2 clinical isolates and reference strains (B-339 and Pb 01 like) were submitted to SDS- Polyacrilamide gel electrophoresis (SDS-PAGE). DNA from clinical isolates and reference strains were submitted to Polymerase chain reaction (PCR) to analyses Internally Transcribed Spacer (ITS) regions 1 and 2. P. brasiliensis strains exhibited great variability of the morphological aspects, when compared with B-339 and Pb 01 like strains. Yeast cells of Pb WAL and Pb1744 presented spherical cells, surrounded by oval and elongated ones, with high incidence of budding yeast cells. While, it was possible to observe small pseudo-hyphae and hyphae forms only from B 339 strains, even after more than 10 sequential cultures on Yeast-Peptone-Dextrose at 35ºC. The cellular extract of the yeast form produced protein and glycoprotein profile with molecular mass ranging 6 kDa to > 180 kDa. The clinical isolates (Pb WAL and Pb 1744) presented distinct protein profile. Overall, the PCR with ITS1/ITS2 (region ITS 1), ITS3/ITS4 (region ITS 2), ITS1/ITS4 (region ITS 1 and 2) primers amplified 300 bp, 400 bp and ~650 bp DNA fragments in all isolates tested, respectively. These are the first studies with P. brasiliensis isolates from the state of Pará. Our results showed that it is important to develop other studies to characterize these clinical isolates. Since protein and glycoprotein differences we found may implicate in modifications concerning the production of exoantigen for serological tests for the diagnosis of PCM.

Keyword: Morphological characteristics, Paracoccidioides brasiliensis, PCR;, Profile protein, Regional strains

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