| Qualitative and quantitative determination of quorum sensing inhibition in vitro. | |
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MedLine Citation:
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PMID: 21031317 Owner: NLM Status: In-Process |
Abstract/OtherAbstract:
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The formation of biofilms in conjunction with quorum sensing (QS)-regulated expression of virulence by opportunistic pathogens contributes significantly to immune evasion and tolerance to a variety of antimicrobial treatments. The present protocol describes methods to determine the in vitro efficacy of potential quorum sensing inhibitors (QSIs). Work on Pseudomonas aeruginosa has shown that chemical blockage of QS is a promising new antimicrobial strategy. Several live bacterial reporter systems been developed to screen extracts and pure compounds for QSI activity. Here we describe the usage of reporter strains consisting of a lasB-gfp or rhlA-gfp fusion in P. aeruginosa for qualitative and quantitative evaluation of the inhibition of the two major QS pathways, monitored as reduced expression of green fluorescence. By the use of an in vitro flow cell system it is possible to study the QSI activity by monitoring its ability to interfere with the protective functions of bacterial biofilm. For evaluation of the global effects of QSI compounds, we present a protocol for the DNA microarray-based transcriptomics. Using these in vitro methods it is possible to evaluate the potential of various QSI compounds. |
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Authors:
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Tim Holm Jakobsen; Maria van Gennip; Louise Dahl Christensen; Thomas Bjarnsholt; Michael Givskov |
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Publication Detail:
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Type: Journal Article |
Journal Detail:
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Title: Methods in molecular biology (Clifton, N.J.) Volume: 692 ISSN: 1940-6029 ISO Abbreviation: Methods Mol. Biol. Publication Date: 2011 |
Date Detail:
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Created Date: 2010-10-29 Completed Date: - Revised Date: - |
Medline Journal Info:
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Nlm Unique ID: 9214969 Medline TA: Methods Mol Biol Country: United States |
Other Details:
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Languages: eng Pagination: 253-63 Citation Subset: IM |
Affiliation:
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Department of International Health, Immunology, and Microbiology, Faculty of Health Sciences, University of Copenhagen, Copenhagen, Denmark. |
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From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine
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