XXI ALAM
Resumo:1404-2


Poster (Painel)
1404-2Identification of resistance mechanisms in sequential clinical isolates of Cryptococcus neoformans
Autores:Suélen Andreia Rossi (FCFAR - UNESP - Faculdade de Ciências Farmacêuticas de Araraquara) ; Vanessa Gonçalves Wolf (FCFAR - UNESP - Faculdade de Ciências Farmacêuticas de Araraquara) ; Fernanda Campos Medeiros (FCFAR - UNESP - Faculdade de Ciências Farmacêuticas de Araraquara) ; Haroldo César de Oliveira (FCFAR - UNESP - Faculdade de Ciências Farmacêuticas de Araraquara) ; Fernanda Sangalli Leite (FCFAR - UNESP - Faculdade de Ciências Farmacêuticas de Araraquara) ; Tatiane Benaducci (FCFAR - UNESP - Faculdade de Ciências Farmacêuticas de Araraquara) ; Fernanda Patrícia Gullo (FCFAR - UNESP - Faculdade de Ciências Farmacêuticas de Araraquara) ; Janaina de Cassia Orlandi Sardi (FCFAR - UNESP - Faculdade de Ciências Farmacêuticas de Araraquara) ; Roseli Maria Zancopé Oliveira (FIOCRUZ - Fundação Osvaldo Cruz) ; Maria José Soares Mendes Giannini (FCFAR - UNESP - Faculdade de Ciências Farmacêuticas de Araraquara) ; Ana Marisa Fusco Almeida (FCFAR - UNESP - Faculdade de Ciências Farmacêuticas de Araraquara)

Resumo

The growth of opportunistic infections has caused problems not only in diagnosis but also therapy, mainly due to the emergence of microbial resistance. Cryptococcosis is a serious disease that occurs in immunocompetent individuals and particularly in immunocompromised patients. It is a disease caused by species of encapsulated yeasts and can also affect several species of animals. Cryptococcosis is caused mainly by two pathogenic species, Cryptococcus neoformans (serotypes A, D and AD), Cryptococcus gattii (serotypes B and C) and interspecies hybrids between both species. This study aimed to analyze the total proteome differentially expressed between isolates recovered from a patient with recurrent cryptococose progressing to the resistance to fluconazole. The relative quantification of PCR Real Time ERG11 gene of isolates, since these have shown resistance to fluconazole developing in vitro, acquired, possibly during the patient therapy. The isolates were characterized as susceptible (26), intermediate (27) and resistant (30) to fluconazole. The latter with a mutation in the ERG11 gene, identified in previous studies as P479L. Relative quantification of gene ERG 11 was performed using the actin gene as a reference strain ATCC 90012 and C. neoformans as control. The protein extraction was performed and prepared to perform the isoelectric focusing (first dimension) and electrophoretic separation (second dimension). It was observed that among clinical isolates was sequential differential expression of total protein demonstrating that even though clinical isolates with identical genetic profiles. The evolution of resistance to fluconazole may have influenced the expression of total protein from isolates. We also note that expression of the ERG11 gene of clinical isolate 26 was less (six-fold less) in comparison with a standard strain ATCC 90012 sensitive. The isolates 27 and 30 showed the major expression of ERG11 gene (four and eight times over), respectively, when compared with strain sensitive pattern indicating that overexpression of this gene may also contribute to the development of resistance to fluconazole in these sequential clinical isolates.


Palavras-chave:  Cryptococcus neoformans, Resistance, ERG11, Fluconazole