ABSTRACT
The condition of cells in Pseudomonas aeruginosa biofilms was monitored via the electrochemical detection of the electro-active virulence factor pyocyanin in a fabricated microfluidic growth chamber coupled with a disposable three electrode cell. Cells were exposed to 4, 16, and 100 mg L(-1) colistin sulfate after overnight growth. At the end of testing, the measured maximum peak current (and therefore pyocyanin concentration) was reduced by approximately 68% and 82% in P. aeruginosa exposed to 16 and 100 mg L(-1) colistin sulfate, respectively. Samples were removed from the microfluidic chamber, analyzed for viability using staining, and streaked onto culture plates to confirm that the P. aeruginosa cells were affected by the antibiotics. The correlation between electrical signal drop and the viability of P. aeruginosa cells after antibiotic exposure highlights the usefulness of this approach for future low cost antibiotic screening applications.
Subject(s)
Anti-Bacterial Agents , Biofilms/drug effects , Electrochemistry/instrumentation , Microbial Sensitivity Tests/methods , Pyocyanine/chemistry , Colistin/chemistry , Dimethylpolysiloxanes/chemistry , Electrochemistry/methods , Electrodes , Escherichia coli/drug effects , Microbial Sensitivity Tests/instrumentation , Microscopy, Electron, Scanning , Pseudomonas aeruginosa/drug effectsABSTRACT
The ability to quickly detect the presence of pathogenic bacteria in patient samples is of the outmost importance to expedient patient care. Here we report the direct, selective, and sensitive detection of the opportunistic pathogen Pseudomonas aeruginosa, spiked in human whole blood with sodium heparin, urine, sputum, and bronchial lavage samples using unmodified, disposable carbon electrode sensors that detect the presence of pyocyanin, a virulence factor that is unique to this species. Square wave voltammetry scans of biological fluids from healthy individuals spiked with P. aeruginosa showed a clear pyocyanin response within one day of culturing at 37°C. Scans of supernatants taken from cultures of P. aeruginosa, Escherichia coli, Staphylococcus aureus, Staphylococcus epidermis, and Bacillus cereus taken over a span of three days in the potential range from -0.5 to 0 V vs. an Ag/AgCl reference showed no electrochemically detectable molecules with the exception of P. aeruginosa. The results indicate the potential to sensitively and selectively determine the presence of P. aeruginosa in human samples via the electrochemical detection of pyocyanin in less than 5 min, without any sample preparation or separation steps.
