Genetic diversity of<i>Histoplasma capsulatum</i>strains in Brazil

Zancopé‐Oliveira, Rosely Maria; Tavares, Patrícia Morais e Silva; Muniz, Mauro de Medeiros

doi:10.1016/j.femsim.2005.05.018

Cited by 21 publications

(13 citation statements)

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“…This study allowed robust analysis of a set of Brazilian H. capsulatum isolates from different geographic regions, and there were three major clusters (clades BrHc1, BrHc2, and BrHc3), as well as clade BrHc4, in each of the four protein-encoding gene trees examined. These data corroborated our initial findings that demonstrated that the phylogenetic population in Brazil is varied (32). Although the genetic separation among the three major clades is considerable, certain isolates exhibited geographic clonality, suggesting that there is partial clonal spread of H. capsulatum within Brazil.…”

Section: Discussionsupporting

confidence: 89%

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Comparison of Different DNA-Based Methods for Molecular Typing of Histoplasma capsulatum

Muniz

Tavares

Meyer

et al. 2010

Appl Environ Microbiol

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Histoplasma capsulatum is very prevalent in the environment and is one of the most common causes of mycoses in humans and diverse animals in Brazil. Multiple typing methods have been developed to study H. capsulatum epidemiology; however, there is limited information concerning comparisons of results obtained with different methods using the same set of isolates. To explore the diversity of H. capsulatum in Brazil and to determine correlations between the results of three different molecular typing techniques, we examined 51 environmental, animal, and human isolates by M13 PCR fingerprinting, PCR-restriction fragment length polymorphism (RFLP) analysis of the internal transcribed region 1 (ITS1)-5.8S-ITS2 region of the rDNA locus, and DNA sequencing and phylogenetic analysis of parts of four protein-encoding genes, the Arf (ADP ribosylation factor), H-anti (H antigen precursor), Ole (delta-9 fatty acid desaturase), and Tub1 (alpha-tubulin) genes. Each method identified three major genetic clusters, and there was a high level of concordance between the results of the typing techniques. The M13 PCR fingerprinting and PCR-RFLP analyses produced very similar results and separated the H. capsulatum isolates included in this study into three major groups. An additional approach used was comparison of our Brazilian ITS1-5.8S-ITS2 sequences with the sequences deposited previously in NCBI data banks. Our analyses suggest that H. capsulatum can be divided into different molecular types that are dispersed around the world. Our results indicate that the three methods used in this study are reliable and reproducible and that they have similar sensitivities. However, M13 PCR fingerprinting has some advantages over the other two methods as it is faster, cheaper, and more user friendly, which especially increases its utility for molecular typing of Histoplasma in situations where laboratory facilities are relatively limited.

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Section: Discussionsupporting

confidence: 89%

“…We reported previously that RAPD analysis was a useful tool for identifying molecular types of H. capsulatum in different geographic regions of Brazil (32). However, we show here that a different PCR-based typing method, M13 PCR fingerprinting, revealed high levels of polymorphism for isolates in a specific region.…”

Section: Discussioncontrasting

confidence: 53%

Comparison of Different DNA-Based Methods for Molecular Typing of Histoplasma capsulatum

Muniz

Tavares

Meyer

et al. 2010

Appl Environ Microbiol

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“…capsulatum isolates were assessed by the RAPD analysis. RAPD reactions were performed with the 31 strains of H. capsulatum using four primers: primer 2 (59-GTTTCGCTCC-39), primer 3 (59-GTAGACCCGT-39), primer 4 (59-AAGAGCCCGT-39) and primer 5 (59-AACGCGCAAC-39), as described by Zancopé-Oliveira et al (2005), with modifications. The RAPD reaction was performed in a total volume of 10 ml, containing 50 ng H. capsulatum DNA, 16 buffer, 1 mM MgCl 2 , 2 pmol primer, 0.5 mM each dNTPs and 1 U Hot Start Taq polymerase (Promega).…”

Section: Methodsmentioning

confidence: 99%

“…capsulatum strains in the southern and south-eastern regions of Brazil (Zancopé-Oliveira et al, 2005;Muniz et al, 2010). However, there is a scarcity of data regarding genotypes circulating in other regions of the country, including the north-east.…”

Section: Introductionmentioning

confidence: 99%

“…capsulatum strains can be grouped according to their geographical origin. According to the study by Zancopé-Oliveira et al (2005), there are three clusters, relating to the north-east (cluster I; n53), south and south-east (cluster II; n517) and midwest regions (cluster III; n52) of Brazil. In this study, the cluster of strains in the north-east region was composed of only three strains from this region, two from Pernambuco and one from Ceará.…”

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Evaluation of the genetic diversity of Histoplasma capsulatum var. capsulatum isolates from north-eastern Brazil

Brilhante

Ribeiro

Lima

et al. 2012

Journal of Medical Microbiology

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Since the beginning of the HIV epidemic, there has been a significant increase in the number of histoplasmosis cases in Ceará , a state in north-east Brazil. The lack of epidemiological data on the genotypes circulating in the north-east region shows the importance of more detailed studies on the molecular epidemiology of Histoplasma capsulatum var. capsulatum in this region. Different molecular techniques have been used to better characterize the genetic profile of H. capsulatum var. capsulatum strains. The aim of this study was to analyse the genetic diversity of H. capsulatum var. capsulatum isolates in Fortaleza, the capital of Ceará , through the sequencing of the internal transcribed spacer (ITS)1-5.8S-ITS2 region, and establish the molecular profile of these isolates, along with strains from south-east Brazil, by RAPD analysis, featuring the different clusters in those regions. The isolates were grouped into two clusters. Cluster 1 included strains from the south-east and north-east regions with separation of isolates into three distinct subgroups (subgroups 1a, 1b and 1c). Cluster 2 included only samples from north-east Brazil. Sequencing of the ITS1-5.8S-ITS2 region allowed the detection of two major clades, which showed geographical correlation between them and their subgroups. Therefore, it can be concluded that the H. capsulatum var. capsulatum isolates from Ceará have a high degree of genetic polymorphism. The molecular data also confirm that populations of this fungus are composed of different genotypes in Brazil and worldwide. INTRODUCTIONHistoplasmosis is an important systemic mycosis, whose aetiological agent is the dimorphic fungus Histoplasma capsulatum. The natural habitat of this fungus is soil, where it grows as a filamentous saprophyte (Kauffman, 2009). This mycosis is widely distributed in the Americas, with a predominance of cases in some areas of the United States, such as along the valleys of the Mississippi and Ohio rivers, as well as in countries of Central and South America (Rossini & Goulart, 2006;Ferreira & Borges, 2009).In Brazil, the occurrence of histoplasmosis is relatively common, as demonstrated by the observation of autochthonous clinical cases, either in isolated cases or in the Abbreviations: ITS, internal transcribed spacer; UPGMA, unweighted pair group analysis with arithmetic mean.The GenBank/EMBL/DDBJ accession numbers for the ITS1-5.8S-ITS2 region sequences of strains used in this study are given in Table 1. Extraction of genomic DNA. Genomic DNA was extracted from yeast-phase cultures of the micro-organism grown for 5 days at 35 uC in Sabouraud agar (Himedia) supplemented with 10 % sheep's blood. DNA extraction was performed as described by Ausubel et al. (2002). Genomic DNA was extracted by mechanical lysis with glass beads and chemistry lysis using a breaking buffer. The nucleic acids were precipitated with ethanol after exposure to a solution of phenol/ chloroform/isoamyl alcohol to separate the cellular debris. The DNA pellet was then resuspended in TE buffer an...

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Histoplasmosis

2011

Fundamental Medical Mycology

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Genetic diversity ofHistoplasma capsulatumstrains in Brazil

Cited by 21 publications

References 15 publications

Comparison of Different DNA-Based Methods for Molecular Typing of Histoplasma capsulatum

Comparison of Different DNA-Based Methods for Molecular Typing of Histoplasma capsulatum

Evaluation of the genetic diversity of Histoplasma capsulatum var. capsulatum isolates from north-eastern Brazil

Histoplasmosis

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