Source: ACCESS SENSOR TECHNOLOGIES LLC submitted to
A LOW-COST PATHOGEN DETECTION SYSTEM FOR FOOD SAFETY
Sponsoring Institution
National Institute of Food and Agriculture
Project Status
TERMINATED
Funding Source
Reporting Frequency
Annual
Accession No.
1006221
Grant No.
2015-33610-23590
Project No.
COLW-2015-00796
Proposal No.
2015-00796
Multistate No.
(N/A)
Program Code
8.5
Project Start Date
Jun 1, 2015
Project End Date
Dec 31, 2016
Grant Year
2015
Project Director
Reilly, T. H.
Recipient Organization
ACCESS SENSOR TECHNOLOGIES LLC
430 N COLLEGE AVE STE 410
FORT COLLINS,CO 80524
Performing Department
(N/A)
Non Technical Summary
Food safety is a global problem, which costs the world billions of dollars and over a million lives annually. Access Sensor Technologies proposes to develop a rapid, low-cost, easy to use system for the measurement of food borne pathogens. The proposed kit builds off of patent-pending technology that will deliver exceptional ease-of-use as well as performance metrics (time, cost, labor) exceeding the current gold standard methods (traditional culture and PCR). By delivering a complete pathogen detection solution, the kit will enable more testing locations and a higher frequency of testing. In addition, the kit is designed to be low-cost from the beginning so entire new segments of food processors may adopt the technology where currently it is not economically feasible to do so, such as rural fruit and vegetable producers that operate only seasonally. Commercial applications of the technology will find a global customer base as current laboratory microbiology tests are cumbersome, expensive and time consuming.The technology proposed by AST will support several of the USDA strategic goals. Low-cost pathogen detection enables American agricultural success by reducing food safety risk which is a central business risk to small co-operatives and independent rural operators that may be unable to survive the crippling financial impact of a single food borne illness outbreak originating from their operations. Economic stability is the result of driving down risks across all areas of business (e.g. finance risk, operations risk, personnel risk, market risk, climate risk). The ultimate goal of a low-cost product that can ensure greater food safety supports USDA strategic goals one, three, and for as outlined in the USDA Strategic Plan.
Animal Health Component
0%
Research Effort Categories
Basic
10%
Applied
20%
Developmental
70%
Classification

Knowledge Area (KA)Subject of Investigation (SOI)Field of Science (FOS)Percent
7125010200080%
7125010110020%
Goals / Objectives
Our goals for this phase I project are:1) Improve detection limits of the paper-based assay.2) Verify Stability and Accuracy of PAD detection.3) Design and Prototype Initial Incubator/AnalyzerOur objectives supporting our goals for this project are:Objective 1: Enrichment Growth Media - Time, exclusivity/Inclusivity AnalysisTask 1.Optimization of L. monocytogenes and S. enterica assays with available growth media.We will test commercial enrichment broths for L. monocytogenes and S. enterica customized to fir our assay. Metrics of evaluation are: enrichment times, exclusivity and inclusivity analysis for the pathogens of interest and pathogens likely to interfere.Task 2: Validation of assays for a given food sample.Here we will test the enrichment broth against a few specific spiked food samples to determine transferability to real world samples.Objective 2: Paper analytical device development and optimizationTask 1. Design and fabrication of PADs for detection of L. monocytogenes and S. enterica.Paper based tests with reagents pre-loaded on paper will be developed. Preparation method, detection limits, reproducibility, geometries and form factors will be determined.Task 2. Reagent stability testing.It is a known concern that some of the molecular compounds used for bacterial pathogen detection have stability/shelf-life issues. Here we will use accelerated shelf-life testing and probe water, oxygen and temperature factors to determine the best strategy for extending and quantifying the shelf life of the paper-based devices.Task 3: Optimize spot size to minimize reagent consumption.Chemical reagent cost is a major consideration for the ultimate product. Here maximizing detection response per unit of reagent used per test will be investigated through geometric and physical variations of the test designs.Task 4: Improve response of PADs with multi-chromogen substrate incorporation.A novel approach to improving detection limits was proposed here that will explore the overlap of multiple colorimetric reagents fixed on the paper based devices. Optical absorbance and reflectance measurements will be carried out to determine if this path can lead to lower detection limits without adding complexity to manufacturing or reading results.Task 5: Validation of methods against molecular detection systems.The improved assay will be compared against gold standard methods.Objective 3: Hardware Design and PrototypeThe paper-based technology will require some support from equipment that will perform timing, heating, cell disruption, and paper incubation. A rough breadboard prototype build from off-the-shelf components along with a form and fit prototype housing cut will be produced to show stakeholders and serve as the starting point for phase II productization activties.
Project Methods
The general scientific methods to be employed for this work are encompassed by the techniques found in microbiology, bio-analytical chemistry, analytical chemistry, materials science, mechanical engineering and electrical engineering.These efforts will be evaluated for success based on the criteria set forth in the proposal. Collecting information from the target audiences will inform and help prioritize success parameters. Determination of overall success and the success of individual tasks will happen through regular internal meetings and data review sessions. Results will be shared with external stakeholders to ensure research and development is moving toward a valuable desired product. These activities include teleconferencing and meeting in-person with our sub-awardees on this grant.

Progress 06/01/15 to 12/31/16

Outputs
Target Audience:The primary audiences are food safety practitioners that may eventually adopt the product under development. These people may be microbiologists by training. In addition stakeholders in the food supply chain are also candidates who benefit from awareness of this product under development. Changes/Problems: Nothing Reported What opportunities for training and professional development has the project provided?As part of this project a graduate student was supported through the sub-award. The company hired a Summer intern who worked for the majority of the Summer supporting the mechanical, electrical and software design of the proposed instrument. How have the results been disseminated to communities of interest?We have continued to meet with potential end-users of the technology to solicit feedback on the product concept. What do you plan to do during the next reporting period to accomplish the goals? Nothing Reported

Impacts
What was accomplished under these goals? Access Sensor Technologies has worked to develop a rapid, low-cost, easy to use system for the measurement of food borne pathogens. The instrument and consumables will deliver exceptional ease-of-use as well as strong performance metrics (time, cost, labor savings). By delivering a complete pathogen detection system we will enable more testing locations and a higher frequency of testing. In addition, the system is designed to be low-cost from the beginning so entire new segments of food processors may adopt the technology where currently it is not economically feasible to do so, such as rural fruit and vegetable producers that operate seasonally. We expect the products will find a global customer base as current laboratory microbiology tests are cumbersome, expensive and time consuming. The technology under development by AST will support several of the USDA strategic goals. Low-cost pathogen detection enables American agricultural success by reducing food safety risk which is a central business risk to small co-operatives and independent rural operators. Economic stability is the result of driving down risks across all areas of business (e.g. finance risk, operations risk, personnel risk, market risk, climate risk). The successes demonstrated thus far show we have reduced technical product risk and confirmed strong interest from the private sector that our product vision has a substantial market opportunity. Our primary goals for the phase I project were to reduce the technical risks associated with our low-cost pathogen detection system. We set out to improve detection limits of the paper-based assay, verify the stability and accuracy of our low-cost consumable devices and design and prototype initial incubator/analyzer. To address our first goal thirteen broths were investigated as potential single-step enrichments for chromogenic detection on PADs. Both selective and non-selective broths were investigated. Many of the broths were able to enrich listeria sufficiently such that when tested at 1 cfu/mL a positive result was detected on our consumable test strip. We also tested the geometries and loadings of reagents on our consumable devices to maximize response and cost effectiveness. To address our second goal we investigated the individual reagents used for our test strip consumables and found that standard storage conditions would be sufficient to support research, manufacturing and inventory storage. We determined test spot geometries that balance readability with reagent economy in order to reduce our projected cost of goods sold. The final significant effort we engaged in was a preliminary design of the prototype instrument that would support our low-cost consumables. We learned quite a bit through interviews with potential end-users about their high priority features. We developed a functional prototype that includes individual reservoirs for incubation broth. Each of these reservoirs may be independently addressed allowing for unique conditions to aid in developing presumptive and confirmatory methods. We also developed preliminary software and analysis algorithms to aid in the automated determination of positive/negative reading of our colorimetric spot tests.

Publications


    Progress 06/01/15 to 05/31/16

    Outputs
    Target Audience:The primary audiences are food safety practitioners that may eventually adopt the product under development. These people may be microbiologists by training. In addition stakeholders in the food supply chain are also candidates who benefit from awareness of this product under development. Changes/Problems: Nothing Reported What opportunities for training and professional development has the project provided?The sub-award for the project has provided training to a graduate student in the field of microbiology. In addition the company created a part-time job for a bachelor level scientist to learn analytical techniques related to colorimetric pathogen detection. How have the results been disseminated to communities of interest?The company developed several realtionships with professionals in the food industry to gauge interest and input on the product under development. As part of these activities we raised awareness of teh USDA SBIR program and its benefits to the food industry. What do you plan to do during the next reporting period to accomplish the goals?With the remaing funds we intend to finish our phase I goals to reduce any remaining technical risks identified during phase I. We will carry out some additional shelf-life testing of assembled products and make continued progress toward a fully functional prototype instrument.

    Impacts
    What was accomplished under these goals? Access Sensor Technologies has worked to develop a rapid, low-cost, easy to use system for the measurement of food borne pathogens. The instrument and consumables will deliver exceptional ease-of-use as well as strong performance metrics (time, cost, labor savings). By delivering a complete pathogen detection system we will enable more testing locations and a higher frequency of testing. In addition, the system is designed to be low-cost from the beginning so entire new segments of food processors may adopt the technology where currently it is not economically feasible to do so, such as rural fruit and vegetable producers that operate seasonally. We expect the products will find a global customer base as current laboratory microbiology tests are cumbersome, expensive and time consuming. The technology under development by AST will support several of the USDA strategic goals. Low-cost pathogen detection enables American agricultural success by reducing food safety risk which is a central business risk to small co-operatives and independent rural operators. Economic stability is the result of driving down risks across all areas of business (e.g. finance risk, operations risk, personnel risk, market risk, climate risk). The successes demonstrated thus far show we have reduced technical product risk and confirmed strong interest from the private sector that our product vision has a substantial market opportunity. Our primary goals for the phase I project were to reduce the technical risks associated with our low-cost pathogen detection system. We set out to improve detection limits of the paper-based assay, verify the stability and accuracy of our low-cost consumable devices and design and prototype initial incubator/analyzer. To address our first goal thirteen broths were investigated as potential single-step enrichments for chromogenic detection on PADs. Both selective and non-selective broths were investigated. Many of the broths were able to enrich listeria sufficiently such that when tested at 1 cfu/mL a positive result was detected on our consumable test strip. We also tested the geometries and loadings of reagents on our consumable devices to maximize response and cost effectiveness. To address our second goal we investigated the individual reagents used for our test strip consumables and found that standard storage conditions would be sufficient to support research, manufacturing and inventory storage. We determined test spot geometries that balance readability with reagent economy in order to reduce our projected cost of goods sold. The final significant effort we engaged in was a preliminary design of the prototype instrument that would support our low-cost consumables. We learned quite a bit through interviews with potential end-users about their high priority features. We will be incorporating many of these features going forward in phase II that will allow for a semi-automated process to ease the challenge of pathogen testing.

    Publications