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Journal of Clinical Microbiology, April 2004, p. 1450-1459, Vol. 42, No. 4
0095-1137/04/$08.00+0     DOI: 10.1128/JCM.42.4.1450-1459.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.

Dynamics of Long-Term Colonization of Respiratory Tract by Haemophilus influenzae in Cystic Fibrosis Patients Shows a Marked Increase in Hypermutable Strains

Federico Román,1 Rafael Cantón,2 María Pérez-Vázquez,1 Fernando Baquero,2 and José Campos1*

Centro Nacional de Microbiología, Instituto de Salud Carlos III, Majadahonda,1 Department of Microbiology, Hospital Universitario Ramón y Cajal, Madrid, Spain2

Received 31 October 2003/ Returned for modification 5 December 2003/ Accepted 22 December 2003

The persistence and variability of 188 Haemophilus influenzae isolates in respiratory tract of 30 cystic fibrosis (CF) patients over the course of 7 years was studied. Antibiotic susceptibility testing, DNA fingerprinting, and analysis of outer membrane protein profiles were performed on all isolates. A total of 115 distinct pulsed-field gel electrophoresis profiles were identified. Ninety percent of patients were cocolonized with two or more clones over the studied period. A third of the patients were cross-colonized with one or two H. influenzae strains; 11% of the clones persisted for 3 or more months. Biotype, outer membrane protein profiles, and resistance profiles showed variation along the studied period, even in persisting clones. Four isolates (2.1%) recovered from 3 patients were type f capsulate, with three of them belonging to the same clone. ß-Lactamase production was detected in 23.9% of isolates while 7% of the ß-lactamase-negative isolates presented diminished susceptibility to ampicillin (ß-lactamase-negative ampicillin resistance phenotype). Remarkably, 21.3% of the H. influenzae isolates presented decreased susceptibility to ciprofloxacin, which was mainly observed in persisting clones. Of the H. influenzae isolates from CF patients, 18 (14.5%) were found to be hypermutable in comparison with 1 (1.4%) from non-CF patients (P < 0.0001). Ten patients (33.3%) were colonized by hypermutable strains over the study period. A multiresistance phenotype and long-term clonal persistence were significantly associated in some cases for up to 7 years. These results suggest that H. influenzae bronchial colonization in CF patients is a dynamic process, but better-adapted clones can persist for long periods of time.

* Corresponding author. Mailing address: Centro Nacional de Microbiología, Instituto de Salud Carlos III, Carretera de Pozuelo, 28220 Majadahonda, Madrid, Spain. Phone: 34-91-822-3650. Fax: 34-91-509-7966. E-mail: jcampos{at}isciii.es.

Journal of Clinical Microbiology, April 2004, p. 1450-1459, Vol. 42, No. 4
0095-1137/04/$08.00+0     DOI: 10.1128/JCM.42.4.1450-1459.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.



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