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PNA-Based Fluorescence In Situ Hybridization for Identification of Bacteria in Clinical Samples

  • Mustafa Fazli
  • Thomas Bjarnsholt
  • Niels Høiby
  • Michael Givskov
  • Tim Tolker-NielsenEmail author
Part of the Methods in Molecular Biology book series (MIMB, volume 1211)

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.

Key words

PNA-FISH Biofilm infections Chronic wounds Cystic fibrosis Soft tissue fillers 

Notes

Acknowledgement

This work was supported by grants from the Lundbeck Foundation, the Danish Strategic Research Council, and the Danish Council for Independent Research.

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Copyright information

© Springer Science+Business Media New York 2014

Authors and Affiliations

  • Mustafa Fazli
    • 1
    • 2
  • Thomas Bjarnsholt
    • 1
    • 3
  • Niels Høiby
    • 1
    • 3
  • Michael Givskov
    • 1
    • 4
  • Tim Tolker-Nielsen
    • 1
    Email author
  1. 1.Costerton Biofilm Center, Department of International Health, Immunology and Microbiology, Faculty of Health and Medical SciencesUniversity of CopenhagenCopenhagenDenmark
  2. 2.Department of Biology, Faculty of ScienceUniversity of CopenhagenCopenhagenDenmark
  3. 3.Department of Clinical MicrobiologyRigshospitaletCopenhagenDenmark
  4. 4.Singapore Centre on Environmental Life Sciences Engineering (SCELSE)Nanyang Technological UniversitySingaporeSingapore

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