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The goal of this project is to develop an inexpensive and robust toxin and pathogen detection system for the rapid (i.e., seconds to minutes), capable of the simultaneous and automated detection of as many as 100 different foodborne toxins and pathogens in meat and poultry processing lines, while promoting interdisciplinary training for graduate and undergraduate students in the fields of optical spectroscopy, multi-spectral imaging, biosensing and chemometric analyses. Our primary research objective is to make fundamental and revolutionary improvements over the current state-of-the-art in the detection of unknown bacteria and/or toxins on and within food products (i.e., chicken, beef, etc.) by: (1) developing novel, inexpensive nano-structured biosensor arrays that work in conjunction with paramagnetic immuno-nanoparticles for the rapid (i.e., seconds to minutes) detection and identification of multiple bacterial and toxic species simultaneously in complex matrices, and (2) employing optically amplified signals, for trace species detection. For this reason, we propose to combine a rapid antibody based pre-concentration and isolation step with a multiplexed SERS-based nanoarray for the specific monitoring of as many as 100 different species in a single measurement. This approach will allow multiple pathogens and toxins to be detected, while providing the desired high assay sensitivities and low false positive results essential for "real-time" food quality monitoring.
NON-TECHNICAL SUMMARY: Foods tainted with various foodborne pathogens and bacteria can result in severe illness in people who consume them. This project is aimed at developing and evaluating a novel multiplexed detection system for the simultaneous monitoring of multiple foodborne pathogenic species.
<P>APPROACH: This goal will be achieved by developing, testing, and demonstrating the efficacy of novel, surface enhanced Raman scattering (SERS)-based nano-biosensor arrays for the redundant monitoring of trace quantities of multiple pathogenic and toxic substances from raw meat and poultry products. The unique nano-photonic properties of these ordered nanosensor arrays (i.e., electromagnetic field enhancement, tunable excitation, etc.) will enable the rapid detection and identification of pathogenic and/or toxic substances in food samples, while providing minimal false positive results due to the redundant biosensing elements. This rapid and accurate diagnostic will result in an increase in the number of quality assurance spot tests that can be performed, as compared to current techniques, resulting in a significant decrease in the shipping of contaminated poultry and meat, without a reduction in production and processing rates. We propose a comprehensive, bottom-up approach toward the real-time monitoring and identification of trace quantities of multiple toxic and pathogenic substances in meat and poultry products. In the proposed research, coherent SERS nano-biosensing arrays with 30,000 individual sensing elements will be constructed, evaluated and optimized for the monitoring of foodborne pathogens. To demonstrate the potential of this SERS nanoarray assay system for the rapid, multiplexed detection of foodborne bacteria and toxins, we will develop and demonstrate the potential of a prototype system for the simultaneous detection of multiple pathogen and toxin simulants in "real-world" samples. This demonstration will be accomplished through a series of tasks/milestones over the 36-month period of this program. These tasks will be accomplished in three phases: (1) Fabrication and optimization of the SERS nanoarrays and complementary capture nanospheres, (2) Development and evaluation of the multi-spectral optical detection system for multiplexed analyses and (3) Validation and testing of the novel SERS nanosphere-nanoarray sensing system for the rapid monitoring of bacteria and toxins on foods.