title: Genotyping, serotyping and determination of mating-type of Cryptococcus neoformans clinical isolates from São Paulo State, Brazil.
abstract: The basidiomycetous yeast Cryptococcus neoformans is an important fungal pathogen mainly in immunocompromised patients. In this study, 47 clinical isolates of C. neoformans from regions of São Paulo State were studied serologically by using the Crypto Check Iatron RM 304-K kit, their genetic diversity was estimated by PCR-fingerprinting with a microsatellite-specific sequence (GACA)(4), RAPD with primer 6 (Amersham Pharmacia Biotech), PCR-restriction fragment length polymorphism (RFLP) analysis of the phospholipase B gene (PLB1) digested with AvaI and mating type analysis by PCR. All 47 strains isolated from HIV positive patients included in this study were serotype A and MATalpha. The majority of the isolates (45/47) were VNI and only two were VNII by PCR-fingerprinting and PCR-RFLP analysis. High degree of homogeneity was observed when (GACA)(4) was used, being highly correlated (> 0.9). In contrast, the RAPD analysis was more heterogeneous with higher number of molecular profiles. By PCR-RFLP, no new molecular type was found, enhancing the suggestion that the differences based on conserved gene as PLB1, can be resultant of ongoing divergent evolution within the C. neoformans complex, into the current eight subtypes. Our results furnish new information on the molecular epidemiology of C. neoformans in the southeast region of Brazil.
authors: Departamento de Análises Clínicas, Faculdade de Ciências Farmacêuticas, Universidade Estadual Paulista, Araraquara, SP, Brasil.
authors: Marcelo Teruyuki
authors: MT
authors: Matsumoto
authors: Ana Marisa
authors: AM
authors: Fusco-Almeida
authors: Lilian Cristiane
authors: LC
authors: Baeza
authors: Márcia de Souza Carvalho
authors: Mde S
authors: Melhem
authors: Maria José Soares
authors: MJ
authors: Medes-Giannini
mesh_terms: D017088:AIDS-Related Opportunistic Infections / Q000382:microbiology; D001938:Brazil; D003455:Cryptococcus neoformans / Q000145:classification / Q000235:genetics* / Q000302:isolation & purification; D016172:DNA Fingerprinting; D049770:Genes, Mating Type, Fungal; D014644:Genetic Variation; D005838:Genotype; D006801:Humans; D016533:Mycological Typing Techniques / Q000379:methods; D012150:Polymorphism, Restriction Fragment Length; D019105:Random Amplified Polymorphic DNA Technique; D012703:Serotyping