4 March 2010 Detection of food-borne pathogens by nanoparticle technology coupled to a low-cost cell reader
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Abstract
The detection, identification and quantification of pathogenic microorganisms at low cost are of great interest to the agro-food industry. We have developed a simple, rapid, sensitive, and specific method for detection of food-borne pathogens based on use of nanoparticles alongside a low cost fluorescence cell reader for the bioassay. The nanoparticles are coupled with antibodies that allow specific recognition of the targeted Listeria in either a liquid or food matrix. The bioconjugated nanoparticles (FNP) contain thousands of dye molecules enabling significant amplification of the fluorescent signal emitted from each bacterium. The developed fluorescence Cell Reader is an LED-based reader coupled with suitable optics and a camera that acquires high resolution images. The dedicated algorithm allowed the counting of each individual nanoparticles-fluorescent bacterial cells thus enabling highly sensitive reading. The system allows, within 1 hour, the recovery and counting of 104 to 108 cfu/mL of Listeria in pure culture. However, neither the Cell Reader nor the algorithm can differentiate between the FNPs specifically-bound to the target and the residual unbound FNPs limiting sensitivity of the system. Since FNPs are too small to be washed in the bioassay, a dual tagging approach was implemented to allow online optical separation of the fluorescent background caused by free FNPs.
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Isabelle Noiseux, Jean-Pierre Bouchard, Pascal Gallant, Pascal Bourqui, Honghe Cao, Marci Vernon, Roger Johnson, Shu Chen, Ozzy Mermut, "Detection of food-borne pathogens by nanoparticle technology coupled to a low-cost cell reader", Proc. SPIE 7553, Frontiers in Pathogen Detection: From Nanosensors to Systems, 75530C (4 March 2010); doi: 10.1117/12.842424; https://doi.org/10.1117/12.842424
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