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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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
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