Use of 16S rRNA gene profiling by terminal restriction fragment length polymorphism analysis to compare bacterial communities in sputum and mouthwash samples from patients with cystic fibrosis

Abstract

The bacterial communities present in the oral cavity and the lungs of 19 adult cystic fibrosis (CF) patients were compared by using terminal restriction fragment length polymorphism analysis of 16S rRNA gene PCR products amplified from nucleic acids extracted directly from bacteria in clinical samples. Sputum samples were not found to be subject to profound contamination by oral cavity bacteria. Evidence of colonization of the CF lung by certain oral bacterial species was found.

FIG. 1.

T-RFLP profiles generated from sputum and mouthwash samples taken from six patients in the sample group. Arrows labeled a, b, and c correspond to the position of bands generated for Haemophilus influenzae, Staphylococcus aureus, and Pseudomonas aeruginosa, respectively.

FIG. 2.

Dendrogram constructed using hierarchical cluster analysis (Dice measure) showing the relative similarities of T-RFLP profiles generated from the sample set.