Please use this identifier to cite or link to this item: https://www.arca.fiocruz.br/handle/icict/32206
Title: Comparison of different DNA-based methods for molecular typing of Histoplasma capsulatum
Authors: Muniz, Mauro de Medeiros
Tavares, Patrícia Morais e Silva
Meyer, Wieland
Nosanchuk, Joshua Daniel
Zancope-Oliveira, Rosely Maria
Affilliation: Fundação Oswaldo Cruz. Instituto de Pesquisa Clínica Evandro Chagas. Laboratório de Micologia. Rio de Janeiro, RJ, Brasil.
Fundação Oswaldo Cruz. Instituto de Pesquisa Clínica Evandro Chagas. Laboratório de Micologia. Rio de Janeiro, RJ, Brasil.
Westmead Millennium Institute. Centre for Infectious Diseases and Microbiology. Molecular Mycology Research Laboratory. New South Wales, Australia / Westmead Hospital. New South Wales, Australia.
Albert Einstein College of Medicine. Microbiology and Immunology. New York, NY, USA.
Fundação Oswaldo Cruz. Instituto de Pesquisa Clínica Evandro Chagas. Laboratório de Micologia. Rio de Janeiro, RJ, Brasil.
Abstract: 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.
Keywords: Histoplasma capsulatum
Molecular typing techniques
Issue Date: 2010
Publisher: American Society for Microbiology
Citation: MUNIZ, Mauro de Medeiros et al. Comparison of different DNA-based methods for molecular typing of Histoplasma capsulatum. Applied and Environmental Microbiology, v. 76, n. 13, p. 4438-4447, July 2010.
DOI: 10.1128/AEM.02004-09
ISSN: 0099-2240
Copyright: open access
Appears in Collections:INI - Artigos de Periódicos

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