Alhede, Morten

Bjarnsholt, Thomas

Høiby, Niels

Johansen, Ulla

Nielsen, Per Halkjær

Rudkjøbing, Vibeke Børsholt

Thomsen, Trine Rolighed

English

Johansen, Ulla R.

Detection of microorganisms involved in airway infection of cystic fibrosis patients by standard culturing &amp; molecular methods

Rudkjøbing, VB

Johansen, Ulla R

Copenhagen University Research Information System

Detection of microorganisms involved in airway infection of cystic fibrosis patients by standard culturing 6 molecular methods

    Aalborg UniversitetDetection of microorganisms involved in airway infection of cystic fibrosis patients bystandard culturing  and molecular methodsRudkjøbing, Vibeke Børsholt; Thomsen, Trine Rolighed; Nielsen, Per Halkjær; Alhede,Morten; Johansen, Ulla; Høiby, Niels; Bjarnsholt, ThomasPublication date:2010Document VersionEarly version, also known as pre-printLink to publication from Aalborg UniversityCitation for published version (APA):Rudkjøbing, V. B., Thomsen, T. R., Nielsen, P. H., Alhede, M., Johansen, U., Høiby, N., & Bjarnsholt, T. (2010).Detection of microorganisms involved in airway infection of cystic fibrosis patients by standard culturing andmolecular methods. Poster presented at The 24th annual north American Cystic Fibrosis Conference 2010,Baltimore, Maryland, United States.General rightsCopyright and moral rights for the publications made accessible in the public portal are retained by the authors and/or other copyright ownersand it is a condition of accessing publications that users recognise and abide by the legal requirements associated with these rights.            ? Users may download and print one copy of any publication from the public portal for the purpose of private study or research.            ? You may not further distribute the material or use it for any profit-making activity or commercial gain            ? You may freely distribute the URL identifying the publication in the public portal ?Take down policyIf you believe that this document breaches copyright please contact us at vbn@aub.aau.dk providing details, and we will remove access tothe work immediately and investigate your claim.Downloaded from vbn.aau.dk on: November 29, 2020  The 24th annual north American cystic fibrosis conference, 2010, Baltimore, Maryland  Poster: Detection of microorganisms involved in airway infection of cystic fibrosis patients by standard culturing and molecular methods.  Vibeke B. Rudkjøbing1, Trine R. Thomsen1,2, Per H. Nielsen1, Morten Alhede3, Ulla R Johansen, Niels Høiby4 and Thomas Bjarnsholt4,5*  The lungs of chronically infected Cystic fibrosis (CF) patients have been intensively studied. Pseudomonas aeruginosa is reported to be the predominant pathogen in CF (Baurnfeind et al. 1987, Koch 2002), however lately a debate of the true bacterial load has occurred. The diagnostics for CF rely mainly on culture based techniques performed on expectorated sputum samples, and most studies are centered on this sample type. Using molecular methods researchers have detected many more bacteria, especially many anaerobes in sputum and BAL samples. It is however problematic since it is possible that the samples may have been contaminated by oral flora during expectoration and BAL sampling. We have in a recent publication only detected P. aeruginosa by PNA FISH (Bjarnsholt 2009) in tissue samples of explanted CF lungs. To study the true bacterial load in CF patients at the Copenhagen CF Center we investigated direct sampled material from explanted lungs (n=4) of CF patients by both culturing and PCR. 

Detection of microorganisms involved in airway infection of cystic fibrosis patients by standard culturing and molecular methods

    Aalborg UniversitetDetection of microorganisms involved in airway infection of cystic fibrosis patients bystandard culturing and molecular methodsRudkjøbing, Vibeke Børsholt; Nielsen, Per Halkjær; Johansen, Ulla; Høiby, Niels; Thomsen,Trine Rolighed; Bjarnsholt, ThomasPublication date:2010Document VersionEarly version, also known as pre-printLink to publication from Aalborg UniversityCitation for published version (APA):Rudkjøbing, V. B., Nielsen, P. H., Johansen, U., Høiby, N., Thomsen, T. R., & Bjarnsholt, T. (2010). Detection ofmicroorganisms involved in airway infection of cystic fibrosis patients by standard culturing and molecularmethods. Abstract from Biofilm IV International Conference, 1-3 september 2010, Winchester, United Kingdom.General rightsCopyright and moral rights for the publications made accessible in the public portal are retained by the authors and/or other copyright ownersand it is a condition of accessing publications that users recognise and abide by the legal requirements associated with these rights.            ? Users may download and print one copy of any publication from the public portal for the purpose of private study or research.            ? You may not further distribute the material or use it for any profit-making activity or commercial gain            ? You may freely distribute the URL identifying the publication in the public portal ?Take down policyIf you believe that this document breaches copyright please contact us at vbn@aub.aau.dk providing details, and we will remove access tothe work immediately and investigate your claim.Downloaded from vbn.aau.dk on: November 29, 2020   Biofilms4 international conference. 1-3 September, 2010. Winchester, UK  Poster: Detection of microorganisms involved in airway infection of cystic fibrosis patients by cultivation techniques and molecular methods. Vibeke B Rudkjøbing1, Trine R Thomsen1,2, Per H Nielsen1, Morten Alhede3, Ulla R Johansen4, Niels Høiby4 and Thomas Bjarnsholt4,5  Patients suffering from the genetic disease cystic fibrosis (CF), develop chronic lung infection. This infection persists due to highly viscous mucus occurring as a result of the disease, in which bacteria form biofilms. Diagnostic tools for CF rely on culture based techniques performed on expectorated sputum samples, and most studies are centered on this sample type. It is however problematic to investigate biofilm-residing microorganisms, and further problems may occur since it is possible that the samples may be contaminated by oral flora during expectoration.  In this study tissue and sputum samples (n=24) from explanted lungs of four Danish CF patients were examined to circumvent possible oral flora contamination. Samples were examined by standard culturing techniques, including aerobic and anaerobic growth, at Rigshospitalet, Denmark. These findings were compared to results obtained by 16S rRNA gene analysis (16S rRNA gene amplification, cloning, sequencing and phylogenetic analysis) performed blinded of the growth results and quantification of the oprL gene of Pseudomonas aeruginosa by quantitative PCR at Aalborg University, Denmark.  The microorganism detected most often by 16S rRNA gene analysis in a sample was also detected by standard culturing techniques (which gave monomicrobial results). 16S rRNA gene analysis suggested that samples contained polymicrobial infection. A correspondence between the frequent detection of P. aeruginosa by the above methods and the number of P. aeruginosa cells in most samples were found by quantitative PCR. This suggests that detection of P. aeruginosa by culturing and 16S rRNA gene analysis is not caused by biases in the techniques. 

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Detection of microorganisms involved in airway infection of cystic fibrosis patients by standard culturing and molecular methods

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