10-minute assay for detecting Escherichia coli O157:H7 in ground beef samples using piezoelectric-excited millimeter-size cantilever sensors.
We detected Escherichia coli O157:H7 (EC) at approximately 10 cells per ml in spiked ground beef samples in 10 min using piezoelectric-excited millimeter-size cantilever (PEMC) sensors. The composite PEMC sensors have a sensing area of 2 mm2 and are prepared by immobilizing a polyclonal antibody specific to EC on the sensing surface. Ground beef (2.5 g) was spiked with EC at 10 to 10,000 cells per ml in phosphate-buffered saline (PBS). One milliliter of supernatant was removed from the blended samples and used to perform the detection experiments. The total resonant frequency change obtained for the inoculated samples was 138 +/- 9, 735 +/- 23, 2,603 +/- 51, and 7,184 +/- 606 Hz, corresponding to EC concentrations of 10, 100, 1,000, and 10,000 cells per ml, respectively. EC was detected in the sample solution within the first 10 min. The responses of the sensor to positive, negative, and buffer controls were 36 +/- 6, 27 +/- 2, and 2 +/- 7 Hz, respectively. Verification of EC attachment was confirmed by low-pH buffer release (PBS-HCl, pH 2.2), microscopy, and second antibody EC binding postdetection. The results indicate that PEMC sensors can reliably detect EC at less than 10 cells per ml in 10 min without sample preparation and with label-free reagents.
Maraldo
David
D
Department of Chemical Engineering, Drexel University, Philadelphia, Pennsylvania 19104, USA.
Mutharasan
Raj
R
D000818:Animals; D015374:Biosensing Techniques / Q000295:instrumentation / Q000379:methods*; D002417:Cattle; D015169:Colony Count, Microbial / Q000379:methods*; D003257:Consumer Product Safety; D019453:Escherichia coli O157 / Q000302:isolation & purification*; D005506:Food Contamination / Q000032:analysis*; D005516:Food Microbiology; D006801:Humans; D008461:Meat Products / Q000382:microbiology*; D013997:Time Factors