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Doctoral Thesis
DOI
Document
Author
Full name
Tânia Sueli de Andrade
Institute/School/College
Knowledge Area
Date of Defense
Published
São Paulo, 2004
Supervisor
Committee
Cury, Arlete Emily (President)
Cunha, Paulo Rowilson
Gambale, Valderez
Mamizuka, Elsa Masae
Paula, Claudete Rodrigues
Title in Portuguese
Caracterização fenotípica e genotípica de cepas de Fonsecaea pedrosoi isoladas de pacientes com cromoblastomicose
Keywords in Portuguese
Cromoblastomicose (Diagnóstico)
Fonsecaea pedrosoi
Fungos (Características)
Micologia médica
PCR
RAPD
RFLP
Abstract in Portuguese
A cromoblastomicose é uma infecção subcutânea causada por fungos demácios, como a Fonsecaea pedrosoi. O pleomorfismo e a similaridade entre gêneros e espécies, ocorrências características dessa classe de fungos, podem interferir na identificação morfológica. Vários esquemas terapêuticos para o tratamento da cromoblastomicose já foram utilizados, mas até o momento não existe uma terapia padrão. Em alguns casos, a correta identificação do agente etiológico pode auxiliar na indicação terapêutica, pois algumas espécies respondem melhor a determinados tratamentos do que outras. Com o objetivo de auxiliar o diagnóstico laboratorial da cromoblastomicose, foi desenvolvido um teste para identificação molecular e detecção rápida da Fonsecaea pedrosoi, empregando-se a metodologia da PCR-duplex com iniciadores específicos para a F. pedrosoi e universais para fungos. Esse teste mostrou-se altamente específico para a F. pedrosoi e, portanto, indicado para ser utilizado na identificação molecular desse fungo. Também com o intuito de avaliar as características fenotípicas e genotípicas de cepas de F. pedrosoi oriundas de pacientes com vários anos de infecção, 47 amostras, isoladas de modo seqüencial, de 11 pacientes foram analisadas por ERIC-PCR, REP-PCR, RFLP e sequenciamento da região ITS1-5.8S-ITS2 do complexo rDNA. Por ERIC-PCR e REP-PCR foram detectados 15 genótipos e, por RFLP, 6 genótipos. As técnicas combinadas geraram 20 genótipos. Com base nos resultados de sequenciamento, foi possível detectar dois tipos de seqüência que apresentaram 97% de similaridade sendo homólogas as seqüências de Fonsecaea pedrosoi disponíveis em bancos gênicos. De modo geral, pode-se observar que a espécie F. pedrosoi apresenta variação molecular e genética entre diferentes cepas e mesmo entre as cepas isoladas de um mesmo paciente.
Title in English
Phenotypic and genotypic characterization of Fonsecaea pedrosoi strains isolated from patients with chromoblastomycosis
Keywords in English
Cromoblastomicose (Diagnóstico)
Fonsecaea pedrosoi
Fungos (Características)
Micologia médica
PCR
RAPD
RFLP
Abstract in English
The chromoblastomycosis is a subcutaneous infection caused by dematiaceous fungi, such as the Fonsecaea pedrosoi. The pleomorfism and the similarity between genus and species that are characteristics of these fungi, can interfere on the morphologic identification. Several therapeutic regimens for the chromoblastomycosis treatment have already been used, but tell now there is not any standard therapy. In some cases, the correct identification of the etiologic agent can help in the treatment because some species have a better response to some treatments than to others. A test was developed to the molecular identification and quick detection of the Fonsecaea pedrosoi with the purpose of helping the laboratorial diagnoses of the chromoblastomycosis, using the PCR - duplex methodology with specific oligonucleotide primers for the Fonsecaea pedrosoi and universals for general fungi. This test is highly specific and sensible for the Fonsecaea pedrosoi and so, it is indicated for the molecular identification of this fungus. Another purpose of this study was to evaluate phenotypic and genotypic characteristics of the Fonsecaea pedrosoi strains isolated from patients with a long-term infection. For this reason 47 samples, isolated sequentially, from 11 patients were analyzed by ERIC-PCR, REP-PCR, RFLP and sequencing of the region ITS1 - 5.8S - ITS2 of the rDNA complex. Fifteen genotypes were found by ERIC-PCR and REP-PCR and six genotypes were found by RFLP, the combination of the methods showed twenty genotypes. Based on the results of sequencing it was possible to detect 2 types sequences that presented 97% of similarity and the available Fonsecaea pedrosoi sequences in gene banks were homological. In general, the Fonsecaea pedrosoi strains presents a molecular and genetic variation between different strains and even between isolated strains of the same patients.
 
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Publishing Date
2019-10-18
 
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