Navegando por Palavras-chave "Micologia Médica"

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    Avaliação e comparação de métodos laboratoriais para diagnóstico das onicomicoses – clássico, molecular e MALDI-TOF
    (Universidade Federal de São Paulo (UNIFESP), 2020-12-01) Santana, Gilglecia Novaes Pereira [UNIFESP]; Camargo, Zoilo Pires De [UNIFESP]; Universidade Federal de São Paulo
    Onychomycosis or fungal nail infection is the infection of the nail bed caused by dermatophytes, yeasts or non dermatophyte filamentous fungi. The infection is not considered serious, although it represents a public health issue due to its high prevalence and associated morbidity, which negatively influences the quality of life of patients. The general objective of this study was to evaluate laboratory methods for the precise and specific diagnosis of the etiologic agents of onychomycosis in immunocompetent (IC) and immunocompromised patients (renal transplant receptors- RTR) with clinical suspicion of onychomycosis. seen at the Dermatology Department of the Federal University of São Paulo (UNIFESP / EPM), Dermatology Service of the Heliópolis Hospital / São Paulo and Private Dermatological Office / São Paulo. Participants were included in this study only after signing the informed consent form. The clinical material of the nails was collected from 50 IC and 50 RTR patients. The isolates were identified by phenotypic, molecular methods and mass spectrometry (MALDI-TOF MS). The susceptibility profile was carried out using the broth microdilution method against the drugs amorolfine, isoconazole, terbinafine itraconazole, fluconazole. The prevalence of onychomycosis was 80% in the IC group and lowered in the RTR group with 76%. In CI patients, the prevalence of onychomycosis was found in females in 72.5%, while in the RTR group, the prevalence was in males in 78.95%. In our investigation, onychomycosis was caused mainly by dermatophytes (55.18%), followed by yeasts 24.14%, FFND and NF were similar, both with 10.34% in the total group in both groups, the T. rubrum species complex was the most frequent, followed by T. mentagrophytes variety T. interdigitale. Regarding yeasts in our investigation, there was a predominance of the genus Candida among the isolates, with the complex of species C. parapsilosis prevalent in both groups. However, in our study, the presence of FFND was found only in the IC group, where no immunodeficiency was observed. As for the identification by mass spectrometry (MALDI-TOF MS), the on-line MSI-IHEM platform, produced significantly better results compared to the Biotyper Bruker database. Based on the high identification rate and the low percentage of errors, our study suggests that the on-line MSI platform is a promising method for the identification of fungi. Despite the statistical similarity between phenotypic and molecular techniques and mass spectrometry (MSI-IHEM Platform), our results reinforce that the molecular technique is more accurate, sensitive to the level of the species complex, providing correct identification, with precision and no error, even without using specific markers, this makes it the gold standard method in our study and has also been reported by several authors. Our data also revealed that the and susceptibility of the isolated species is similar to those described in the literature.
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    Taxonomia polifásica e características proteômicas do complexo Sporothrix schenckii
    (Universidade Federal de São Paulo (UNIFESP), 2010-08-25) Rodrigues, Anderson Messias [UNIFESP]; Camargo, Zoilo Pires de [UNIFESP]; Universidade Federal de São Paulo (UNIFESP)
    Sporotrichosis is a chronic mycotic infectious disease of man and animals caused by the traumatic implantation of a pathogenic fungus that typically involves the skin and subcutaneous tissue. Since 1898, when the sporotrichosis etiological agent was discovered by Schenck, this disease has been attributed to a single pathogen, Sporothrix schenckii Hektoen & Perkins, a thermo dimorphic fungus that grow as a yeast at 37 ºC and as a mycelium at room temperature. However, isolates identified as S. schenckii showed a great deal of genetic variability, suggesting that this taxon consist in a cryptic species complex. Based on this information our group is interested in the study of the polyphasic taxonomy of this species complex and its implication on its ecoepidemiology in Brazil. We studied 161 strains of Sporothrix spp. provided from environmental and clinical samples obtained from diverse regions of Brazil and other countries. The phenotypic parameters assayed include vegetative growth on PDA media at different temperatures (30, 35, 37 and 40 ºC), the colony colors on CMA media, the assimilation pattern of carbon sources (raffinose, ribitol and sucrose) and morphological microscopic features of in vitro cultivation. The phenotypic data were confirmed by molecular biology using the sequencing information of a fragment of calmodulin locus. Our data show that the S. brasiliensis, S. globosa, S. mexicana and S. schenckii have a widespread geographical distribution in Brazil. To our knowledgement this is the first description of the specie S. mexicana outside of Mexico and causing disease in human host. S. brasiliensis was isolated with high frequency from cats in Rio Grande do Sul and Rio de Janeiro states, suggesting that cats has been an important vector in epidemiology of this specie. A proteomic approach was proposed in this work to compare protein profile of Sporothrix spp. isolates and get a better understanding about the differences in terms of protein expression among the cryptic species. The 2-D profiles were strongly different among the isolates. To elucidate the major antigen of human sporotrichosis, the fungus was cultured in BHI broth, 37 °C, and intracellular proteins were resolved by 2-DE. Proteins were transferred to a nitrocellulose membrane and then incubated with serum from patients with the two major clinical form of disease (cutaneous fixed and linfocutaneous). Our results show that IgG present in serum from patients react with different antigens from Sporothrix schenckii complex. Immunoblotting showed that the sera of patients had antibodies reacting with a 70 kDa antigen in three isolates (S. brasiliensis, S. globosa and S. schenckii). Profile differences in antigenicity were observed between S. mexicana and the other species studied here.