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

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Book cover In Situ Hybridization Protocols

Part of the book series: Methods in Molecular Biology ((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.

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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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Authors and Affiliations

  1. Costerton Biofilm Center, Department of International Health, Immunology and Microbiology, Faculty of Health and Medical Sciences, University of Copenhagen, Blegdamsvej 3B, 2200, Copenhagen, Denmark

    Mustafa Fazli, Thomas Bjarnsholt, Niels Høiby, Michael Givskov & Tim Tolker-Nielsen

  2. Department of Biology, Faculty of Science, University of Copenhagen, 2200, Copenhagen, Denmark

    Mustafa Fazli

  3. Department of Clinical Microbiology, Rigshospitalet, 2100, Copenhagen, Denmark

    Thomas Bjarnsholt & Niels Høiby

  4. Singapore Centre on Environmental Life Sciences Engineering (SCELSE), Nanyang Technological University, Singapore, Singapore

    Michael Givskov

Authors
  1. Mustafa Fazli
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  2. Thomas Bjarnsholt
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  3. Niels Høiby
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  4. Michael Givskov
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  5. Tim Tolker-Nielsen
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Corresponding author

Correspondence to Tim Tolker-Nielsen .

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Editors and Affiliations

  1. Molecular Histology, Bioneer A/S, Horsholm, Denmark

    Boye Schnack Nielsen

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© 2014 Springer Science+Business Media New York

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Fazli, M., Bjarnsholt, T., Høiby, N., Givskov, M., Tolker-Nielsen, T. (2014). PNA-Based Fluorescence In Situ Hybridization for Identification of Bacteria in Clinical Samples. In: Nielsen, B. (eds) In Situ Hybridization Protocols. Methods in Molecular Biology, vol 1211. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-1459-3_21

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  • DOI: https://doi.org/10.1007/978-1-4939-1459-3_21

  • Published:

  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-1458-6

  • Online ISBN: 978-1-4939-1459-3

  • eBook Packages: Springer Protocols

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