Comparison of the sensitivity of culture, PCR and quantitative real-time PCR for the detection of Pseudomonas aeruginosa in sputum of cystic fibrosis patients |
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| Authors: | Pieter Deschaght Thierry De Baere Leen Van Simaey Sabine Van daele Frans De Baets Daniel De Vos Jean-Paul Pirnay Mario Vaneechoutte |
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| Institution: | (1) Laboratory for Bacteriology Research (LBR), Ghent University Hospital, University of Ghent, Ghent, Belgium;(2) Cystic Fibrosis Centre, Department Pediatric Pulmonology, Ghent University Hospital, University of Ghent, Ghent, Belgium;(3) Laboratory for Molecular and Cellular Technology (LabMCT), Burn Wound Center, Queen Astrid Military Hospital, Brussels, Belgium |
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| Abstract: | Background
Pseudomonas aeruginosa is the major pathogen involved in the decline of lung function in cystic fibrosis (CF) patients. Early aggressive antibiotic
therapy has been shown to be effective in preventing chronic colonization. Therefore, early detection is important and sensitive
detection methods are warranted. In this study, we used a dilution series of P. aeruginosa positive sputa, diluted in a pool of P. aeruginosa negative sputa, all from CF patients - to mimick as closely as possible the sputa sent to routine laboratories - to compare
the sensitivity of three culture techniques versus that of two conventional PCR formats and four real-time PCR formats, each
targeting the P. aeruginosa oprL gene. In addition, we compared five DNA-extraction protocols. |
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