ANTIFUNGAL SUSCEPTIBILITY TESTING AND GENOTYPING CHARACTERIZATION OF Cryptococcus neoformans AND gattii ISOLATES FROM HIV-INFECTED PATIENTS OF RIBEIRÃO PRETO, SÃO PAULO, BRAZIL.
Cryptococcosis is a leading invasive fungal infection in immunocompromised patients. Considering the high prevalence and severity of these infections in immunocompromised patients attended at HC-FMRP-USP, the present research aimed to characterize the clinical isolates of Cryptococcus strains by biochemical and molecular methods and evaluate antifungal susceptibility of clinical isolates. Fifty isolates from 32 HIV-positive patients were obtained at HC-FMRP-USP. Most of the isolates (78.1%) were identified as C. neoformans, and 100% of C. neoformans and C. gattii strains were susceptible to amphotericin B, ketoconazole and fluconazole. All isolates were classified as serotype A (grubbii variety) by PCR and most of them were characterized in mating type MATa. PCR analysis of specific M13 microsatellite sequence revealed that VNI type was predominant among C. neoformans, while VGII was predominant among C. gattii. The strains did not show a significant resistance to the antifungals tested, and Canavanine-Glycine-Bromthymol Blue Agar (CGB) proved to be a reliable test presenting a good correlation with the molecular characterization. C. neoformans isolated from disseminated infections in the same patient showed molecular identity when different anatomical sites were compared; besides, the studied strains did not present a significant increase in resistance to antifungal agents. In addition, the homogeneity of the molecular types and detection of the mating types suggested a low possibility of crossing among the strains.
Universidade de São Paulo, Faculdade de Medicina de Ribeirão Preto, Departamento de Clínica Médica. 14049-900 Ribeirão Preto, SP, Brazil. E-mail: thais.pandini@gmail.com.|Universidade Federal de Ouro Preto, Núcleo de Inovação Tecnológica, 35400-000 Ouro Preto, MG, Brazil. E-mail: thais.pandini@gmail.com.
Thais Pandini
TP
Figueiredo
Universidade de São Paulo, Faculdade de Medicina de Ribeirão Preto, Departamento de Bioquímica. 14049-900 Ribeirão Preto, SP, Brazil. E-mail: rosymar_lucas@hotmail.com.|Universidade de São Paulo, Faculdade de Filosofia, Ciências e Letras de Ribeirão Preto, Departamento de Biologia. 14040-901 Ribeirão Preto, SP, Brazil. E-mail: rosymar_lucas@hotmail.com.
Rosymar Coutinho de
RC
Lucas
Universidade Federal de Sergipe, Departamento de Morfologia. 49100-000, SE, Brazil. E-mail: cazzaniga.rodrigo@gmail.com.
Rodrigo Anselmo
RA
Cazzaniga
Universidade de Santo Amaro, UNISA, Pós Graduação em Ciências da Saúde, São Paulo, SP, Brazil. E-mail: carolufscar24@gmail.com.|Universidade Federal de São Paulo, UNIFESP, Departamento de Medicina, São Paulo, SP, Brazil. E-mail: carolufscar24@gmail.com.
Carolina Nunes
CN
França
, Escola de Engenharia de Lorena, Departamento de Biotecnologia. 12602-810, Lorena, SP,Universidade de São Paulo Brazil. E-mail: segatof@icloud.com.
Fernando
F
Segato
Universidade de São Paulo, Faculdade de Medicina de Ribeirão Preto, Departamento de Biologia Celular, Molecular e Bioagentes Patogênicos. 14049-900 Ribeirão Preto, SP, Brazil. E-mails: rafaeltaglialegna@hotmail.com; cmlmaffe@fmrp.usp.br.
Rafael
R
Taglialegna
Universidade de São Paulo, Faculdade de Medicina de Ribeirão Preto, Departamento de Biologia Celular, Molecular e Bioagentes Patogênicos. 14049-900 Ribeirão Preto, SP, Brazil. E-mails: rafaeltaglialegna@hotmail.com; cmlmaffe@fmrp.usp.br.
Claudia Maria Leite
CM
Maffei
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