Performing Department
(N/A)
Non Technical Summary
Approximately 47.8 million episodes (one in six Americans) of foodborne illnesses happen each year and result in roughly 128,000 hospitalizations and 3,000 deaths in the U.S. Approximately half of the reported foodborne illnesses occur in children, with the majority occurring in children under 15years of age. The development of reliable and effective methods to detect foodborne hazards (pathogens, microorganisms, chemicals, toxins) is therefore of paramount importance. To address the USDA's need for a field-ready device to rapidly detect foodborne hazards during pre- and post-harvest processing and distribution, Physical Optics Corporation (POC) proposes to develop a novel rapid Foodborne Pathogen Detection (FPAD) system based on POC's established sandwich ELISA and lab-on-a-chip technologies. The system can perform ELISA with reagent volumes <1-mL and <60 min assay time, leading to a significant reduction of costs and technician-hours associated with time-consuming lab testing. The new system design allows for quantitative identification, agnostic sample preparation, process automation beneficial to users with little training, multiplex assay capability, and high portability. In Phase I, POC will demonstrate FPAD feasibility by assembling a benchtop prototype and conducting multiplex immunoassays for detection of three selected pathogens. In Phase II, POC plans to further develop a fully operational FPAD system by enhancing accuracy, reducing costs, increasing ruggedness for use under extreme field environments, improving configurability for customizable assays (including pathogens and biological toxins), and implementing system automation.With POC's FPAD, the nation will have a cost-effective and efficient platform for rapid identification of microbial pathogens and biological toxins from food, water, and other environmental samples, which is beneficial to federal agencies, such as the Centers for Disease Control and Prevention (CDC). Other applications include on-site detection of pathogens during foodborne illness events, use by First Responders for onsite analysis of harmful pathogens in unknown samples, human health, veterinary applications, and food contamination. Its pathogen-detection capability will also be useful in point-of-care diagnostics for bacterial infections in human and animal subjects, screening for environmental toxins, and assessment of the efficacy of sanitary and decontamination procedures.
Animal Health Component
0%
Research Effort Categories
Basic
10%
Applied
45%
Developmental
45%
Goals / Objectives
Approximately 47.8 million episodes (one in six Americans) of foodborne illnesses happen each year and result in roughly 128,000 hospitalizations and 3,000 deaths in the U.S. The major goal of this project is to develop a reliable, field-ready device to rapidly detect foodborne hazards (pathogens, microorganisms, chemicals, toxins) during pre- and post-harvest processing, distribution, and storage. In Phase I, POC will demonstrate FPAD feasibility by assembling a benchtop prototype and conducting multiplex immunoassays for detection of three selected pathogens.Objective 1. Design and development of the FPAD system.Objective 2. Assembly and preliminary laboratory testing of the FPAD prototype.Objective 3. Feasibility demonstration of the FPAD prototype for efficient pathogen identification.Objective 4. Definition of the commercial market for FPAD technology.
Project Methods
The project will be conducted based on the following tasks. Tasks 5 and 6 will be performed to evaluate the FPAD prototype.Task 1. Design the System Architecture of the FPAD PlatformPOC will design the architecture of the FPAD system, including the dimensions and configuration of the microfluidic cartridge, the volumetric flow velocity of the sample and reagents, the architecture of the optical interrogation unit, and the electronic requirements.Task 2. Develop the AssayPOC will develop the assay to target three pathogens from the top ten pathogens responsible for the vast majority of foodborne illness events in the U.S., as identified by the CDC. We will optimize the assay conditions of sandwich ELISA for multiplex pathogen detection.Task 3. Design and Fabricate the FPAD Microfluidic CartridgePOC will work closely with ALine, Inc., a local microfluidics manufacturing company, to fabricate the FPAD microfluidic chips, which will be cost-effective when mass produced. We will incorporate a glass slide with immobilized capture antibody spots onto a microfluidic chip. Other parts of the cartridge, including sample/reagent reservoirs and pneumatic valves, will be developed and integrated into the microfluidic chip in Phase II.Task 4. Design and Assemble the Optical Interrogation Unit, Microfluidic Control Unit, and Electronic UnitThe optical interrogation unit, microfluidic control unit, and electronic unit of the Phase I FPAD prototype will be developed and assembled. The interrogation unit will be situated on a motorized positioning stage for rapid scan through all the detection spots. The power consumption of each unit will be determined to allow for a Phase I FOBID prototype that can be powered via a universal serial bus (USB) port.Task 5. Assemble the FPAD Prototype and Perform Initial TestingPOC will assemble a proof-of-concept, benchtop FPAD prototype, which will integrate the microfluidic cartridges, microfluidic control unit, optical interrogation unit, and electronic unit, and use a computer to synchronize the immunoassay process and perform data measurement. Initial testing will be conducted to optimize the operating parameters of the components.Task 6. Demonstrate the Feasibility of the FPAD PrototypePOC will conduct laboratory testing and validation of the FPAD prototype by demonstrating the capability of the prototype to quantitatively identify three pathogens from laboratory-prepared samples. The limit of detection, sensitivity, accuracy, and false positive/negative rates will be determined. Software and algorithms will be developed for ELISA process control, interrogation, and data analysis to demonstrate ease of use. In addition, POC will test the FPAD prototype against samples prepared by mixing known pathogens with food samples, such as vegetables and meat, followed by homogenization and filtration for feasibility demonstration.Task 7. Explore Commercial Potential and Product ViabilityPOC will explore the potential to transfer the FPAD technology to the USDA. The cost will be estimated to determine the market competitiveness. Market research will identify the most promising applications of the FPAD system. Sources of Phase II and Phase III guidance and matching funds will be identified early in the project, and these early business partners will be involved throughout development and commercialization.