Molecular epidemiology and phylogenetic analysis of Haemophilus parainfluenzae from chronic obstructive pulmonary disease exacerbations

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Abstract
This study was undertaken to characterize serial Haemophilus parainfluenzae strains from epidemiologically unrelated chronic obstructive pulmonary disease (COPD) patients and from healthy carriers. A comprehensive approach was used including different phenotypical and molecular typing methods: biotyping, antibiotyping, conventional ribotyping, pulsed field gel electrophoresis (PFGE) assay, and PCR-ribotyping. Conventional ribotyping and PFGE analysis were confirmed as excellent procedures to differentiate isolates of the same species and biotype. Conversely, in our study, PCR-ribotyping proved to be suitable for taxonomic purposes, unambiguously identifying H. parainfluenzae from H. influenzae but not discriminating strains at the intraspecific level for epidemiological typing. Phylogenetic analysis of restriction fragment length polymorphism (RFLP) data of sequences related to the rrn operon demonstrated that H. parainfluenzae strains associated to COPD are spread among many diverging lineages.