Genotyping, serotyping and determination of mating-type of Cryptococcus neoformans clinical isolates from São Paulo State, Brazil

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.

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Main Authors: Matsumoto,Marcelo Teruyuki, Fusco-Almeida,Ana Marisa, Baeza,Lilian Cristiane, Melhem,Márcia de Souza Carvalho, Medes-Giannini,Maria José Soares
Format: Digital revista
Language:English
Published: Instituto de Medicina Tropical de São Paulo 2007
Online Access:http://old.scielo.br/scielo.php?script=sci_arttext&pid=S0036-46652007000100008
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spelling oai:scielo:S0036-466520070001000082007-03-19Genotyping, serotyping and determination of mating-type of Cryptococcus neoformans clinical isolates from São Paulo State, BrazilMatsumoto,Marcelo TeruyukiFusco-Almeida,Ana MarisaBaeza,Lilian CristianeMelhem,Márcia de Souza CarvalhoMedes-Giannini,Maria José Soares Cryptococcus Epidemiology Mating type RAPD PCR fingerprinting RFLP 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.info:eu-repo/semantics/openAccessInstituto de Medicina Tropical de São PauloRevista do Instituto de Medicina Tropical de São Paulo v.49 n.1 20072007-02-01info:eu-repo/semantics/articletext/htmlhttp://old.scielo.br/scielo.php?script=sci_arttext&pid=S0036-46652007000100008en10.1590/S0036-46652007000100008
institution SCIELO
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country Brasil
countrycode BR
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libraryname SciELO
language English
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author Matsumoto,Marcelo Teruyuki
Fusco-Almeida,Ana Marisa
Baeza,Lilian Cristiane
Melhem,Márcia de Souza Carvalho
Medes-Giannini,Maria José Soares
spellingShingle Matsumoto,Marcelo Teruyuki
Fusco-Almeida,Ana Marisa
Baeza,Lilian Cristiane
Melhem,Márcia de Souza Carvalho
Medes-Giannini,Maria José Soares
Genotyping, serotyping and determination of mating-type of Cryptococcus neoformans clinical isolates from São Paulo State, Brazil
author_facet Matsumoto,Marcelo Teruyuki
Fusco-Almeida,Ana Marisa
Baeza,Lilian Cristiane
Melhem,Márcia de Souza Carvalho
Medes-Giannini,Maria José Soares
author_sort Matsumoto,Marcelo Teruyuki
title Genotyping, serotyping and determination of mating-type of Cryptococcus neoformans clinical isolates from São Paulo State, Brazil
title_short Genotyping, serotyping and determination of mating-type of Cryptococcus neoformans clinical isolates from São Paulo State, Brazil
title_full Genotyping, serotyping and determination of mating-type of Cryptococcus neoformans clinical isolates from São Paulo State, Brazil
title_fullStr Genotyping, serotyping and determination of mating-type of Cryptococcus neoformans clinical isolates from São Paulo State, Brazil
title_full_unstemmed Genotyping, serotyping and determination of mating-type of Cryptococcus neoformans clinical isolates from São Paulo State, Brazil
title_sort genotyping, serotyping and determination of mating-type of cryptococcus neoformans clinical isolates from são paulo state, brazil
description 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.
publisher Instituto de Medicina Tropical de São Paulo
publishDate 2007
url http://old.scielo.br/scielo.php?script=sci_arttext&pid=S0036-46652007000100008
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