title: PNA-based fluorescence in situ hybridization for identification of bacteria in clinical samples.
abstract: Fluorescence in situ hybridization with PNA probes (PNA-FISH) that target specific bacterial ribosomal RNA sequences is a powerful and rapid tool for identification of bacteria in clinical samples. PNA can diffuse readily through the bacterial cell wall due to its uncharged backbone, and PNA-FISH can be performed with high specificity due to the extraordinary thermal stability of RNA-PNA hybrid complexes. We describe a PNA-FISH procedure and provide examples of the application of PNA-FISH for the identification of bacteria in chronic wounds, cystic fibrosis lungs, and soft tissue fillers. In all these cases, bacteria can be identified in biofilm aggregates, which may explain their recalcitrance to antibiotic treatment.
authors.affiliation: Costerton Biofilm Center, Department of International Health, Immunology and Microbiology, Faculty of Health and Medical Sciences, University of Copenhagen, Blegdamsvej 3B, 2200, Copenhagen, Denmark.
authors.forename: Mustafa
authors.initials: M
authors.lastname: Fazli
authors.forename: Thomas
authors.initials: T
authors.lastname: Bjarnsholt
authors.forename: Niels
authors.initials: N
authors.lastname: Høiby
authors.forename: Michael
authors.initials: M
authors.lastname: Givskov
authors.forename: Tim
authors.initials: T
authors.lastname: Tolker-Nielsen
mesh_terms: D001419:Bacteria / Q000235:genetics / Q000302:isolation & purification*; D018441:Biofilms / Q000254:growth & development; D003550:Cystic Fibrosis / Q000382:microbiology; D016591:Histocytological Preparation Techniques / Q000379:methods; D006801:Humans; D017404:In Situ Hybridization, Fluorescence / Q000379:methods*; D020135:Peptide Nucleic Acids / Q000032:analysis* / Q000235:genetics; D012329:RNA, Bacterial / Q000032:analysis* / Q000235:genetics; D018461:Soft Tissue Infections / Q000382:microbiology; D013183:Sputum / Q000382:microbiology; D014947:Wounds and Injuries / Q000382:microbiology
chemical_list: D020135:Peptide Nucleic Acids; D012329:RNA, Bacterial