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Molecular typing of the bacterial flora in sputum of cystic fibrosis patients
Stockholm Cystic Fibrosis Centre, Huddinge University Hospital, Stockholm, Sweden.
Linköping University, Faculty of Health Sciences. Linköping University, Department of Clinical and Experimental Medicine, Clinical Microbiology . Östergötlands Läns Landsting, Centre for Laboratory Medicine, Department of Molecular Biological Techniques.
Linköping University, Faculty of Health Sciences. Linköping University, Department of Clinical and Experimental Medicine, Molecular and Immunological Pathology . Östergötlands Läns Landsting, Centre for Laboratory Medicine, Department of Clinical Pathology and Clinical Genetics.
2003 (English)In: International Journal of Medical Microbiology, ISSN 1438-4221, E-ISSN 1618-0607, Vol. 293, no 4, 309-317 p.Article in journal (Refereed) Published
Abstract [en]

Despite recent advances in therapy, lower airway infections remain the major cause of morbidity and mortality in cystic fibrosis (CF) patients. Bacterial colonisation of the lower airways in CF is limited to a few bacterial species, commonly Staphylococcus aureus, Pseudomonas aeruginosa and Haemophilus influenzae. Burkholderia cepacia colonisation is much rarer, but it has been thought to be associated with more advanced lung disease and increased mortality. A rapid characterisation of the bacterial flora in sputum of CF patients is of great importance for proper treatment. The aim of this study was to establish bacterial profiles and to identify pathogenic bacteria in respiratory specimens by means of molecular methods including temporal temperature gradient gel electrophoresis (TTGE) and DNA sequencing of PCR amplicons derived from 16S rDNA variable V3 and V6 regions. Sputa of 13 CF patients (7 males/6 females, age 19-59 years) collected at the Stockholm CF centre were analysed. TTGE revealed the presence of complex bacterial profiles in all samples. The V3 and V6 PCR amplicons were cloned and sequenced by real-time DNA PyrosequencingTM. DNA from Staphylococcus aureus, Haemophilus influenzae, and Pseudomonas aeruginosa, respectively, was identified together with sequences from normal oral cavity flora. The results were in reasonable agreement with those obtained by conventional bacterial culture, considering that only known CF pathogens are included in routine reports. However, the methodology seems too elaborate to be introduced into daily routine.

Place, publisher, year, edition, pages
2003. Vol. 293, no 4, 309-317 p.
Keyword [en]
16S rRNA, Bacterial typing, Cystic fibrosis, Pyrosequencing, Sputum, Temporal temperature gradient gel electrophoresis
National Category
Medical and Health Sciences
Identifiers
URN: urn:nbn:se:liu:diva-46534DOI: 10.1078/1438-4221-00265OAI: oai:DiVA.org:liu-46534DiVA: diva2:267430
Available from: 2009-10-11 Created: 2009-10-11 Last updated: 2017-12-13

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Monstein, Hans-JurgJonasson, Jon

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Faculty of Health SciencesClinical Microbiology Department of Molecular Biological TechniquesMolecular and Immunological Pathology Department of Clinical Pathology and Clinical Genetics
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