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In vitro screens for quorum sensing inhibitors and in vivo confirmation of their effect

Nature Protocols volume 5, pages 282–293 (2010)Cite this article

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Abstract

This article will introduce the reader to protocols intended for (i) identification of quorum sensing (QS) inhibitors (QSIs), (ii) characterization of these compounds in vitro and (iii) evaluation of these compounds in animal models. Traditional antimicrobial drugs are designed against planktonic bacteria and not against bacterial biofilms. In biofilms, bacteria are highly resistant to otherwise lethal treatments and they communicate with each other, thus enabling coordinated group behavior. For many years, we have focused on interference with cell to cell communication, also known as QS, with the aim of disabling the expression of virulence and reduction of antibiotic tolerance. Here we present protocols for screening and testing for acyl-homoserine lactone (AHL)-dependent QS inhibition. We also present protocols for the in vivo validation of QSIs as possible drug candidates. The presented methods allow the evaluation of QS inhibition by a potential drug candidate within 2–3 weeks.

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Figure 1: Diagram of the different methods used to identify and determine efficacy of quorum-sensing inhibitors (QSIs).
Figure 2: Setup for bead making.
Figure 3: Quorum sensing inhibitor selector (QSIS) 1 assay.
Figure 4: Example of a potential quorum-sensing inhibitor (QSI) tested over time with the lasB-gfp fusion in Pseudomonas aeruginosa.

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Acknowledgements

We thank Anne Kirstine Nielsen for her contribution to experimental setups.

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Authors and Affiliations

  1. Department of International Health, Immunology and Microbiology, Faculty of Health Sciences, University of Copenhagen, Copenhagen, Denmark

    Thomas Bjarnsholt, Maria van Gennip, Tim H Jakobsen, Louise D Christensen & Michael Givskov

  2. Department of Clinical Microbiology, Rigshospitalet, Copenhagen, Denmark

    Thomas Bjarnsholt & Peter Ø Jensen

Authors
  1. Thomas Bjarnsholt
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  2. Maria van Gennip
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  3. Tim H Jakobsen
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  4. Louise D Christensen
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  5. Peter Ø Jensen
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  6. Michael Givskov
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Contributions

All authors contributed extensively to developing and writing the protocols presented in this paper.

Corresponding author

Correspondence to Michael Givskov.

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Bjarnsholt, T., van Gennip, M., Jakobsen, T. et al. In vitro screens for quorum sensing inhibitors and in vivo confirmation of their effect. Nat Protoc 5, 282–293 (2010). https://doi.org/10.1038/nprot.2009.205

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