Analysis of the Bacterial Communities Present in Lungs of Patients with Cystic Fibrosis from American and British Centers

Molecular Microbiology Research Laboratory, Pharmaceutical Science Research Division, King's College London, 150 Stamford Street, Franklin-Wilkins Building, London, SE1 9NH, United Kingdom.
Journal of clinical microbiology (Impact Factor: 3.99). 11/2010; 49(1):281-91. DOI: 10.1128/JCM.01650-10
Source: PubMed


The aim of this study was to determine whether geographical differences impact the composition of bacterial communities present in the airways of cystic fibrosis (CF) patients attending CF centers in the United States or United Kingdom. Thirty-eight patients were matched on the basis of clinical parameters into 19 pairs comprised of one U.S. and one United Kingdom patient. Analysis was performed to determine what, if any, bacterial correlates could be identified. Two culture-independent strategies were used: terminal restriction fragment length polymorphism (T-RFLP) profiling and 16S rRNA clone sequencing. Overall, 73 different terminal restriction fragment lengths were detected, ranging from 2 to 10 for U.S. and 2 to 15 for United Kingdom patients. The statistical analysis of T-RFLP data indicated that patient pairing was successful and revealed substantial transatlantic similarities in the bacterial communities. A small number of bands was present in the vast majority of patients in both locations, indicating that these are species common to the CF lung. Clone sequence analysis also revealed that a number of species not traditionally associated with the CF lung were present in both sample groups. The species number per sample was similar, but differences in species presence were observed between sample groups. Cluster analysis revealed geographical differences in bacterial presence and relative species abundance. Overall, the U.S. samples showed tighter clustering with each other compared to that of United Kingdom samples, which may reflect the lower diversity detected in the U.S. sample group. The impact of cross-infection and biogeography is considered, and the implications for treating CF lung infections also are discussed.

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    • "As the respiratory microbiome is much more restricted in species diversity as compared to soil or intestinal microbiomes, we used one labeled primer, one restriction nuclease, and the length standard ROX1000 for analysis. Similar approaches were used in related studies using material from patients with cystic fibrosis [35], [36]. The resolution of T-RFLP chromatograms is around 2–5 bp, causing that sometimes separate organisms have similar fragment sizes. "
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