Syracuse

Fluorescence enlightens RND pump activity and the intrabacterial concentration of antibiotics

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Masi, Muriel | Dumont, Estelle | Vergalli, Julia | Pajovic, Jelena | Réfrégiers, Matthieu | Pages, Jean-Marie

Edité par CCSD ; Elsevier -

International audience. To understand the antibiotic resistance in Gram-negative bacteria, a key point is to investigate antibiotic accumulation which is defined by influx and efflux. Several methods exist to evaluate the membrane permeability and efflux pump activity but they present some disadvantages and limitations. An optimized spectrofluorimetric method using the intrinsic tryptophan fluorescence as internal standard as well as a complementary microfluorimetric assay following the time-course accumulation in intact individual cells have been developed. Comparing the latter population and single cell approaches can lead to the understanding of the phenotypic heterogeneity within a population. The two methodologies lead to the determination of parameters, concentration, accumulation rates, localization that contribute to emerging concepts (RTC2T, SICAR) with the aim to identify and detail the antibiotic chemotypes that are involved in influx/efflux.

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