Improved Diagnosis of Biofilm Infections Using Various Molecular Methods
Traditional culture-dependent methods and a number of culture-independent molecular methods including 16S rRNA gene polymerase chain reaction, construction of clone libraries, sequencing, phylogeny, fingerprinting, fluorescence in situ hybridization and quantitative PCR were used to describe the microbial composition of two types of biofilm-related infections, namely chronic venous leg ulcers and prosthetic joint infections. Multiple tissue biopsies were taken from each chronic wound, and different specimen types (joint fluid, tissue biopsy, bone biopsy and prosthesis scraping or sonication) were collected from prosthetic joint patients. The obtained results indicate that in these two types of infections the bacterial composition and yield may vary depending on the position and type of samples used for analysis. It emphasizes the need for multiple samplings in order to achieve better diagnosis and treatment of these biofilm-related infections. The most complete picture of microbial composition of biofilms is probably accomplished when several culture and culture-independent methods are used in parallel to characterize the pathogens.
KeywordsClone Library Molecular Technique Peptide Nucleic Acid Prosthetic Joint Infection qPCR Testing
Thanks to our colleagues in the Danish cross-disciplinary Prosthesis: Reduction of Infection and Pain consortium. The study was supported by a grant from the Danish Agency of Science and Technology (no. 09-052174).
Thomas Bjarnsholt, Bo Jørgensen and Klaus Kirketerp-Møller are acknowledged for great collaboration in the area of chronic ulcers. The Danish Technical Research Council supported this study under the innovation consortia “BIOMED”.
Martin Aasholm, Vibeke Rudkjøbing, Susanne Bielidt and Masumeh Chavoshi are thanked for their valuable technical assistance.
- PRIS website. https://www.knee.dk/groups/grp_login.php. Accessed Aug 2011