Sensing bacterial vibrations and early response to antibiotics with phase noise of a resonant crystal

Ward L. Johnson, Danielle Cook France, Nikki S. Rentz, William T. Cordell, Fred L. Walls

The speed of conventional antimicrobial susceptibility testing (AST) is intrinsically limited by observation of cell colony growth, which can extend over days and allow bacterial infections to advance before effective antibiotics are identified. This report presents an approach for rapidly sensing mechanical fluctuations of bacteria and the effects of antibiotics on these fluctuations. Bacteria are adhered to a quartz crystal resonator in an electronic bridge that is driven by a high-stability frequency source. Mechanical fluctuations of cells introduce time-dependent perturbations to the crystal boundary conditions and associated resonant frequency, which translate into phase noise measured at the output of the bridge. In experiments on nonmotile E. coli exposed to polymyxin B, cell-generated frequency noise dropped close to zero with the first spectra acquired 7 minutes after introduction of the antibiotic. In experiments on the same bacterial strain exposed to ampicillin, frequency noise began decreasing within 15 minutes of antibiotic introduction and proceeded to drop more rapidly with the onset of antibiotic-induced lysis. In conjunction with cell imaging and post-experiment counting of colony-forming units, these results provide evidence that cell death can be sensed through measurements of cell-generated frequency noise, potentially providing a basis for rapid AST.