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Molecular Detection of Multiple Emerging Pathogens in Sputa from Cystic Fibrosis Patients

BACKGROUND: There is strong evidence that culture-based methods detect only a small proportion of bacteria present in the respiratory tracts of cystic fibrosis (CF) patients. METHODOLOGY/PRINCIPAL FINDINGS: Standard microbiological culture and phenotypic identification of bacteria in sputa from CF p...

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Autores principales: Bittar, Fadi, Richet, Hervé, Dubus, Jean-Christophe, Reynaud-Gaubert, Martine, Stremler, Nathalie, Sarles, Jacques, Raoult, Didier, Rolain, Jean-Marc
Formato: Texto
Lenguaje:English
Publicado: Public Library of Science 2008
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2483419/
https://www.ncbi.nlm.nih.gov/pubmed/18682840
http://dx.doi.org/10.1371/journal.pone.0002908
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author Bittar, Fadi
Richet, Hervé
Dubus, Jean-Christophe
Reynaud-Gaubert, Martine
Stremler, Nathalie
Sarles, Jacques
Raoult, Didier
Rolain, Jean-Marc
author_facet Bittar, Fadi
Richet, Hervé
Dubus, Jean-Christophe
Reynaud-Gaubert, Martine
Stremler, Nathalie
Sarles, Jacques
Raoult, Didier
Rolain, Jean-Marc
author_sort Bittar, Fadi
collection PubMed
description BACKGROUND: There is strong evidence that culture-based methods detect only a small proportion of bacteria present in the respiratory tracts of cystic fibrosis (CF) patients. METHODOLOGY/PRINCIPAL FINDINGS: Standard microbiological culture and phenotypic identification of bacteria in sputa from CF patients have been compared to molecular methods by the use of 16S rDNA amplification, cloning and sequencing. Twenty-five sputa from CF patients were cultured that yield 33 isolates (13 species) known to be pathogens during CF. For molecular cloning, 760 clones were sequenced (7.2±3.9 species/sputum), and 53 different bacterial species were identified including 16 species of anaerobes (30%). Discrepancies between culture and molecular data were numerous and demonstrate that accurate identification remains challenging. New or emerging bacteria not or rarely reported in CF patients were detected including Dolosigranulum pigrum, Dialister pneumosintes, and Inquilinus limosus. CONCLUSIONS/SIGNIFICANCE: Our results demonstrate the complex microbial community in sputa from CF patients, especially anaerobic bacteria that are probably an underestimated cause of CF lung pathology. Metagenomic analysis is urgently needed to better understand those complex communities in CF pulmonary infections.
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spelling pubmed-24834192008-08-06 Molecular Detection of Multiple Emerging Pathogens in Sputa from Cystic Fibrosis Patients Bittar, Fadi Richet, Hervé Dubus, Jean-Christophe Reynaud-Gaubert, Martine Stremler, Nathalie Sarles, Jacques Raoult, Didier Rolain, Jean-Marc PLoS One Research Article BACKGROUND: There is strong evidence that culture-based methods detect only a small proportion of bacteria present in the respiratory tracts of cystic fibrosis (CF) patients. METHODOLOGY/PRINCIPAL FINDINGS: Standard microbiological culture and phenotypic identification of bacteria in sputa from CF patients have been compared to molecular methods by the use of 16S rDNA amplification, cloning and sequencing. Twenty-five sputa from CF patients were cultured that yield 33 isolates (13 species) known to be pathogens during CF. For molecular cloning, 760 clones were sequenced (7.2±3.9 species/sputum), and 53 different bacterial species were identified including 16 species of anaerobes (30%). Discrepancies between culture and molecular data were numerous and demonstrate that accurate identification remains challenging. New or emerging bacteria not or rarely reported in CF patients were detected including Dolosigranulum pigrum, Dialister pneumosintes, and Inquilinus limosus. CONCLUSIONS/SIGNIFICANCE: Our results demonstrate the complex microbial community in sputa from CF patients, especially anaerobic bacteria that are probably an underestimated cause of CF lung pathology. Metagenomic analysis is urgently needed to better understand those complex communities in CF pulmonary infections. Public Library of Science 2008-08-06 /pmc/articles/PMC2483419/ /pubmed/18682840 http://dx.doi.org/10.1371/journal.pone.0002908 Text en Bittar et al. http://creativecommons.org/licenses/by/4.0/ This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are properly credited.
spellingShingle Research Article
Bittar, Fadi
Richet, Hervé
Dubus, Jean-Christophe
Reynaud-Gaubert, Martine
Stremler, Nathalie
Sarles, Jacques
Raoult, Didier
Rolain, Jean-Marc
Molecular Detection of Multiple Emerging Pathogens in Sputa from Cystic Fibrosis Patients
title Molecular Detection of Multiple Emerging Pathogens in Sputa from Cystic Fibrosis Patients
title_full Molecular Detection of Multiple Emerging Pathogens in Sputa from Cystic Fibrosis Patients
title_fullStr Molecular Detection of Multiple Emerging Pathogens in Sputa from Cystic Fibrosis Patients
title_full_unstemmed Molecular Detection of Multiple Emerging Pathogens in Sputa from Cystic Fibrosis Patients
title_short Molecular Detection of Multiple Emerging Pathogens in Sputa from Cystic Fibrosis Patients
title_sort molecular detection of multiple emerging pathogens in sputa from cystic fibrosis patients
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2483419/
https://www.ncbi.nlm.nih.gov/pubmed/18682840
http://dx.doi.org/10.1371/journal.pone.0002908
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