19 February 2009 Detection of E. coli O157:H7 in complex matrices under varying flow parameters with a robotic fluorometric assay system
Author Affiliations +
Abstract
The development of rapid assays for detection of microbial pathogens in complex matrices is needed to protect public health due to continued outbreaks of disease from contaminated foods and water. An Escherichia coli O157:H7 detection assay was designed using a robotic, fluorometric assay system. The system integrates optics, fluidics, robotics and software for the detection of foodborne pathogens or toxins in as many as four samples simultaneously. It utilizes disposable fiber optic waveguides coated with biotinylated antibodies for capture of target analytes from complex sample matrices. Computer-controlled rotation of sample cups allows complete contact between the sample and the waveguide. Detection occurs via binding of a fluorophore-labeled antibody to the captured target, which leads to an increase in the fluorescence signal. Assays are completed within twenty-five minutes. Sample matrices included buffer, retentate (material recovered from the filter of the Automated Concentration System (ACS) following hollow fiber ultrafiltration), spinach wash and ground beef. The matrices were spiked with E. coli O157:H7 (103-105 cells/ml) and the limits of detection were determined. The effect of sample rotation on assay sensitivity was also examined. Rotation parameters for each sample matrix included 10 ml with rotation, 5 ml with rotation and 0.1 ml without rotation. Detection occurred at 104 cells/ml in buffer and spinach wash and at 105 cells/ml in retentate and ground beef. Detection was greater for rotated samples in each matrix except ground beef. Enhanced detection of E. coli from large, rotated volumes of complex matrices was confirmed.
© (2009) COPYRIGHT Society of Photo-Optical Instrumentation Engineers (SPIE). Downloading of the abstract is permitted for personal use only.
Stephaney D. Leskinen, Sarah M. Schlemmer, Elizabeth A. Kearns, Daniel V. Lim, "Detection of E. coli O157:H7 in complex matrices under varying flow parameters with a robotic fluorometric assay system", Proc. SPIE 7167, Frontiers in Pathogen Detection: From Nanosensors to Systems, 71670J (19 February 2009); doi: 10.1117/12.807830; https://doi.org/10.1117/12.807830
PROCEEDINGS
10 PAGES


SHARE
RELATED CONTENT

An automated portable array biosensor
Proceedings of SPIE (July 22 2003)
A portable array biosensor for food safety
Proceedings of SPIE (November 19 2004)
Array biosensor: recent developments
Proceedings of SPIE (May 03 1999)
A portable array biosensor
Proceedings of SPIE (December 07 2004)
Autonomous navigation with teams of aerial robots
Proceedings of SPIE (May 13 2011)

Back to Top