Rapid flow cytometric assessment of mecillinam and ampicillin bacterial susceptibility

Abstract

The effects of antibiotics on Escherichia coli during exponential growth in liquid medium were monitored by flow cytometry measuring cellular DNA content and cell size of individual cells in large numbers as well as cell counts. A statistically significant increase in cellular DNA as well as size was recorded after 20 and 40 min of incubation with the MIC of ampicillin and mecillinam, respectively. The optical density (OD(600 nm)) of treated cultures continued to increase for at least 80 minutes, showing that the increase in cellular DNA and size reflected continued growth and elongation of cells coupled with inhibition of cell division, the latter being confirmed by the cell counting. Since fixation, staining and flow-cytometric analysis can be carried out within a few minutes, the present results suggest that flow cytometry may have potential as a rapid and quantitative technique that can be automated for clinical and experimental susceptibility testing of antibacterial drugs.