Comparison of different DNA-based methods for molecular typing of histoplasma capsulatum

Mauro Medeiros De Muniz, Patrícia Morais E Silva Tavares, Wieland Meyer, Joshua D. Nosanchuk, Rosely Maria Zancope-Oliveira

Research output: Contribution to journalArticle

24 Citations (Scopus)

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 (ITSl)-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.

Original languageEnglish (US)
Pages (from-to)4438-4447
Number of pages10
JournalApplied and Environmental Microbiology
Volume76
Issue number13
DOIs
StatePublished - Jul 2010

Fingerprint

Histoplasma capsulatum
Histoplasma
Molecular Typing
DNA
Polymerase Chain Reaction
polymorphism
Restriction Fragment Length Polymorphisms
Brazil
methodology
ADP-Ribosylation Factors
restriction fragment length polymorphism
gene
animal
Complement Factor H
epidemiology
antigen
Mycoses
stearoyl-CoA desaturase
Tubulin
Ribosomal DNA

ASJC Scopus subject areas

  • Applied Microbiology and Biotechnology
  • Food Science
  • Biotechnology
  • Ecology

Cite this

Comparison of different DNA-based methods for molecular typing of histoplasma capsulatum. / De Muniz, Mauro Medeiros; Tavares, Patrícia Morais E Silva; Meyer, Wieland; Nosanchuk, Joshua D.; Zancope-Oliveira, Rosely Maria.

In: Applied and Environmental Microbiology, Vol. 76, No. 13, 07.2010, p. 4438-4447.

Research output: Contribution to journalArticle

De Muniz, Mauro Medeiros ; Tavares, Patrícia Morais E Silva ; Meyer, Wieland ; Nosanchuk, Joshua D. ; Zancope-Oliveira, Rosely Maria. / Comparison of different DNA-based methods for molecular typing of histoplasma capsulatum. In: Applied and Environmental Microbiology. 2010 ; Vol. 76, No. 13. pp. 4438-4447.
@article{920871ebfedc4a7082e95dd1e22cae0f,
title = "Comparison of different DNA-based methods for molecular typing of histoplasma capsulatum",
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 (ITSl)-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.",
author = "{De Muniz}, {Mauro Medeiros} and Tavares, {Patr{\'i}cia Morais E Silva} and Wieland Meyer and Nosanchuk, {Joshua D.} and Zancope-Oliveira, {Rosely Maria}",
year = "2010",
month = "7",
doi = "10.1128/AEM.02004-09",
language = "English (US)",
volume = "76",
pages = "4438--4447",
journal = "Applied and Environmental Microbiology",
issn = "0099-2240",
publisher = "American Society for Microbiology",
number = "13",

}

TY - JOUR

T1 - Comparison of different DNA-based methods for molecular typing of histoplasma capsulatum

AU - De Muniz, Mauro Medeiros

AU - Tavares, Patrícia Morais E Silva

AU - Meyer, Wieland

AU - Nosanchuk, Joshua D.

AU - Zancope-Oliveira, Rosely Maria

PY - 2010/7

Y1 - 2010/7

N2 - 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 (ITSl)-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.

AB - 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 (ITSl)-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.

UR - http://www.scopus.com/inward/record.url?scp=77954303049&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=77954303049&partnerID=8YFLogxK

U2 - 10.1128/AEM.02004-09

DO - 10.1128/AEM.02004-09

M3 - Article

C2 - 20453140

AN - SCOPUS:77954303049

VL - 76

SP - 4438

EP - 4447

JO - Applied and Environmental Microbiology

JF - Applied and Environmental Microbiology

SN - 0099-2240

IS - 13

ER -