Qualitative and Quantitative Determination of Quorum Sensing Inhibition In Vitro
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Qualitative and Quantitative Determination of Quorum Sensing Inhibition In Vitro. / Jakobsen, Tim Holm; Alhede, Maria; Hultqvist, Louise Dahl; Bjarnsholt, Thomas; Givskov, Michael.
Quorum Sensing: Methods and Protocols. Vol. 1673 Springer, 2018. p. 275-285 (Methods in molecular biology (Clifton, N.J.)).Research output: Chapter in Book/Report/Conference proceeding › Book chapter › Research › peer-review
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TY - CHAP
T1 - Qualitative and Quantitative Determination of Quorum Sensing Inhibition In Vitro
AU - Jakobsen, Tim Holm
AU - Alhede, Maria
AU - Hultqvist, Louise Dahl
AU - Bjarnsholt, Thomas
AU - Givskov, Michael
PY - 2018
Y1 - 2018
N2 - 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 QS inhibitors (QSIs). Work on Pseudomonas aeruginosa has shown that chemical blockage of QS is a promising new antimicrobial strategy. Several live bacterial reporter systems have 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 two of the 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.
AB - 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 QS inhibitors (QSIs). Work on Pseudomonas aeruginosa has shown that chemical blockage of QS is a promising new antimicrobial strategy. Several live bacterial reporter systems have 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 two of the 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.
U2 - 10.1007/978-1-4939-7309-5_21
DO - 10.1007/978-1-4939-7309-5_21
M3 - Book chapter
C2 - 29130180
SN - 978-1-4939-7308-8
VL - 1673
T3 - Methods in molecular biology (Clifton, N.J.)
SP - 275
EP - 285
BT - Quorum Sensing
PB - Springer
ER -
ID: 190437007