Analysis of the airway microbiota of healthy individuals and patients with chronic obstructive pulmonary disease by T-RFLP and clone sequencing

PLoS One. 2013 Jul 9;8(7):e68302. doi: 10.1371/journal.pone.0068302. Print 2013.

Abstract

Chronic obstructive pulmonary disease (COPD) is a progressive, inflammatory lung disease that affects a large number of patients and has significant impact. One hallmark of the disease is the presence of bacteria in the lower airways.

Objective: The aim of this study was to analyze the detailed structure of microbial communities found in the lungs of healthy individuals and patients with COPD. Nine COPD patients as compared and 9 healthy individuals underwent flexible bronchoscopy and BAL was performed. Bacterial nucleic acids were subjected to terminal restriction fragment (TRF) length polymorphism and clone library analysis. Overall, we identified 326 T-RFLP band, 159 in patients and 167 in healthy controls. The results of the TRF analysis correlated partly with the data obtained from clone sequencing. Although the results of the sequencing showed high diversity, the genera Prevotella, Sphingomonas, Pseudomonas, Acinetobacter, Fusobacterium, Megasphaera, Veillonella, Staphylococcus, and Streptococcus constituted the major part of the core microbiome found in both groups. A TRF band possibly representing Pseudomonas sp. monoinfection was associated with a reduction of the microbial diversity. Non-cultural methods reveal the complexity of the pulmonary microbiome in healthy individuals and in patients with COPD. Alterations of the microbiome in pulmonary diseases are correlated with disease.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adult
  • Aged
  • Bacteria / classification*
  • Bacteria / genetics*
  • Bronchoalveolar Lavage Fluid / microbiology
  • Case-Control Studies
  • Female
  • Humans
  • Male
  • Microbiota*
  • Middle Aged
  • Molecular Sequence Data
  • Phylogeny
  • Polymorphism, Restriction Fragment Length
  • Pulmonary Disease, Chronic Obstructive / microbiology*
  • RNA, Ribosomal, 16S / genetics
  • Respiratory System / microbiology*
  • Respiratory System / pathology
  • Sequence Analysis, DNA
  • Young Adult

Substances

  • RNA, Ribosomal, 16S

Associated data

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Grants and funding

This study was supported by grants from the Federal Ministry of Education and Research (FKZ 01GI0881-0888) to RB. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.