Affinity-selected filamentous bacteriophage as a probe for acoustic wave biodetectors of Salmonella typhimurium.
Proof-in-concept biosensors were prepared for the rapid detection of Salmonella typhimurium in solution, based on affinity-selected filamentous phage prepared as probes physically adsorbed to piezoelectric transducers. Quantitative deposition studies indicated that approximately 3 x 10(10)phage particles/cm(2) could be irreversibly adsorbed for 1 h at room temperature to prepare working biosensors. The quality of phage deposition was monitored by fluorescent microscopy. Specific-bacterial binding resulted in resonance frequency changes of prepared sensors, which were evaluated using linear regression analysis. Sensors possessed a rapid response time of <180 s, had a low-detection limit of 10(2)cells/ml and were linear over a range of 10(1)-10(7)cells/ml with a sensitivity of 10.9 Hz per order of magnitude of S. typhimurium concentration. Viscosity effects due to increasing bacterial concentration and non-specific binding were not significant to the piezoelectric platform as confirmed by dose-response analysis. Phage-bacterial binding was confirmed by fluorescence and scanning electron microscopy. Overall, phage may constitute effective bioreceptors for use with analytical platforms for detecting and monitoring bacterial agents, including use in food products and possibly biological warfare applications.
Olsen
Eric V
EV
Department of Biological Sciences, Auburn University, 101 Life Sciences Bldg., AL 36849, USA.
Sorokulova
Iryna B
IB
Petrenko
Valery A
VA
Chen
I-Hsuan
IH
Barbaree
James M
JM
Vodyanoy
Vitaly J
VJ
D001435:Bacteriophages / Q000302:isolation & purification* / Q000502:physiology*; D015374:Biosensing Techniques / Q000295:instrumentation* / Q000379:methods; D015169:Colony Count, Microbial / Q000295:instrumentation* / Q000379:methods; D004563:Electrochemistry / Q000295:instrumentation / Q000379:methods; D004867:Equipment Design; D019544:Equipment Failure Analysis; D015336:Molecular Probe Techniques / Q000295:instrumentation*; D015203:Reproducibility of Results; D012486:Salmonella typhimurium / Q000302:isolation & purification* / Q000821:virology*; D012680:Sensitivity and Specificity