Source: PHYSICAL OPTICS CORPORATION submitted to
RAPID FOODBORNE ILLNESS DETECTION SYSTEM
Sponsoring Institution
National Institute of Food and Agriculture
Project Status
COMPLETE
Funding Source
Reporting Frequency
Annual
Accession No.
1006377
Grant No.
2015-33610-23581
Cumulative Award Amt.
$99,999.00
Proposal No.
2015-00785
Multistate No.
(N/A)
Project Start Date
Jun 15, 2015
Project End Date
Feb 14, 2017
Grant Year
2015
Program Code
[8.5]- Food Science & Nutrition
Project Director
Chao, C.
Recipient Organization
PHYSICAL OPTICS CORPORATION
1845 WEST 205TH STREET
TORRANCE,CA 90501
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
45%
Research Effort Categories
Basic
10%
Applied
45%
Developmental
45%
Classification

Knowledge Area (KA)Subject of Investigation (SOI)Field of Science (FOS)Percent
7120499202025%
7120510202075%
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.

Progress 06/15/15 to 02/14/17

Outputs
Target Audience: Nothing Reported Changes/Problems: Nothing Reported What opportunities for training and professional development has the project provided? Nothing Reported How have the results been disseminated to communities of interest? Nothing Reported What do you plan to do during the next reporting period to accomplish the goals? Nothing Reported

Impacts
What was accomplished under these goals? In the course of Phase I, POC accomplished the following: Defined the preliminary operational protocol for sample preprocessing and detection and established the system's operational procedure and architecture to provide rapid (<1 hr) detection of foodborne hazards. The mature FOBID is expected to be very competitive with the current standard approaches (Table 5-1). Developed a Salmonella and Campylobacter assay with a limit of detection (LOD) at 106CFU mL-1 for a 2-µL sample volume. In Phase II, we will focus on the optimization of the preconcentration process and assay to achieve detection of 1 CFU/25 g of foodsample. Designed and fabricated the microfluidic chip with an estimated cost of $75-85 for a screen of ten pathogens in comparison to >$100 using conventional ELISA and PCRapproaches. Designed and assembled the optical interrogation unit and its related electronics and control software for successful demonstration of the FOBID functional capability andfeasibility. Demonstrated the preconcentration process that can be used to process large volumes of food sample. In Phase II, POC will develop this protocol further and build a compact preconcentration module for large volume sampling and LOD reduction. Demonstrated the FOBID's capability to successfully detect and identify Salmonella and Campylobacter antigens in the microfluidic platform, rendering good specificity and low cross reactivity. Explored the commercial potential and product viability of the FOBID system, which has a niche in food safety testing, biodefense-related rapid pathogen detection, and point-of-care diagnostics markets (>$35B in 2021).

Publications