Abstract
The aim of this study was to develop an innovative method of examining bacterial survival using capillary zone electrophoresis (CZE) and flow cytometry (FC) as a reference method. For this purpose, standard strains of bacteria from the ATCC collection were used: Enterococcus faecalis ATCC 14506, Staphylococcus aureus ATCC 11632, Klebsiella pneumoniae ATCC 10031, Pseudomonas aeruginosa ATCC 27853, and Escherichia coli ATCC 25922, as well as seven antibiotics with different antimicrobial mechanisms of action. The ratio of live and dead cells in the tested sample in CZE measurements were calculated using our algorithm that takes into account the detection time. Results showed a high agreement between CZE and FC in the assessment of the percentage of live cells exposed to the stress factor in both antibiotic susceptibility and time-dependent assays. The applied measuring system to assess the effectiveness of antibiotic therapy in in vitro conditions is a method with great potential, and the data obtained with the use of CZE mostly correspond to the expected drug sensitivity according to EUCAST and CLSI guidelines.
Highlights
The bacterial cell is forced to constantly adapt to new changing environmental conditions, such as temperature, pressure, oxygen and carbon dioxide levels, or the presence of water, nutrients or toxic substances
Two different approaches were considered to achieve the aim of our study, which was to determine the suitability of capillary zone electrophoresis in assessing the in vitro efficacy cytometry out using(Enterococcus a MACSQuant
InIn summary, capillary zone electrophoresis appears to be effective in determining the viability of pathogens exposed to substances that change their physiological state
Summary
The bacterial cell is forced to constantly adapt to new changing environmental conditions, such as temperature, pressure, oxygen and carbon dioxide levels, or the presence of water, nutrients or toxic substances. These parameters influence its metabolic activity and change its physiological state [1]. The crucial element in laboratory diagnosis, besides identifying pathogens, is the assessment of their viability and antimicrobial susceptibility testing mostly determine by phenotypic (e.g., minimum inhibitory concentration (MIC) or genotypic approaches, which is essential to understanding the pathogen’s response to therapy [2]. The viability is identified as the ability of a cell to grow and reproduce itself under a set of defined environmental conditions. Viable cells are characterized by the presence and functioning of a range of structural, physiological, metabolic, and genetic properties [3]
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