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Detection and quantification of the heterogeneity of S. aureus bacterial populations to identify antibiotic-induced persistence
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Hassan, Marwa, Butler, Mark S., Ranzoni, Andrea and Cooper, Matthew A. (2018) Detection and quantification of the heterogeneity of S. aureus bacterial populations to identify antibiotic-induced persistence. Working Paper. BioRxiv: Cold Spring Harbour.
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Official URL: http://dx.doi.org/10.1101/320093
Abstract
Objectives: Persister cells are characterised as being viable but non-culturable, a state that preserves their metabolic energy to survive the environmental stress, which allows for recurrent infections. Detection of persisters is, therefore, not possible with standard culture-dependent methods. Furthermore, the effect of antibiotics on the induction of persisters has not been assessed. This study aimed to identify antibiotic-induced persistence and determine the percentage of heterogeneity. Methods: Vancomycin, daptomycin and dalbavancin were assessed by standard MIC methods against selected Staphylococcus aureus strains. Replicates of MIC assays were stained with propidium iodide to quantify live/dead and a reactive oxygen species (ROS) dye to detect and quantify persisters using culture-independent single-cell sorting, independently. A comparative analysis was then performed. Results: Dalbavancin showed the lowest MIC values against tested S. aureus strains followed by daptomycin and vancomycin. Cell sorting of vancomycin-, daptomycin- and dalbavancin-treated S. aureus strains showed a range of 1.9-10.2%, 17.7-62.9% and 7.5-77.6% live cells based on the strain, respectively, in which daptomycin, in particular, was a strong inducer of a persister population. Persisters represented 3.7-16% of the bacterial population. Conclusions: The culture-independent identification of antibiotic-induced persistence through studying at the single-cell level showed different efficacy of antibiotics than standard MIC. Vancomycin was the most effective antibiotic against tested strains followed by dalbavancin then daptomycin as assessed by cell sorting. Therefore, re-evaluation of standard MIC methods may be required to assess the efficacy of antibiotics. Additionally, the detection of daptomycin-associated persisters may provide an elucidation to the reported rapid resistance development in vivo.
Item Type: | Working or Discussion Paper (Working Paper) | ||||
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Divisions: | Faculty of Science, Engineering and Medicine > Science > Life Sciences (2010- ) | ||||
Journal or Publication Title: | BioRxiv | ||||
Publisher: | Cold Spring Harbour | ||||
Place of Publication: | BioRxiv | ||||
Book Title: | Detection and quantification of the heterogeneity of S. aureus bacterial populations to identify antibiotic-induced persistence | ||||
Official Date: | 11 May 2018 | ||||
Dates: |
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Number: | 320093 | ||||
DOI: | 10.1101/320093 | ||||
Institution: | University of Warwick | ||||
Status: | Peer Reviewed | ||||
Publication Status: | Published | ||||
Access rights to Published version: | Restricted or Subscription Access |
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