Source: SMART DIAGNOSTICS SYSTEMS LLC submitted to NRP
RAPID AND PORTABLE OPTICAL DETECTION SYSTEM FOR MULTIPLE FOOD-BORNE PATHOGENS, E. COLI O157:H7 AND SALMONELLA ENTERICA IN MEAT
Sponsoring Institution
National Institute of Food and Agriculture
Project Status
COMPLETE
Funding Source
Reporting Frequency
Annual
Accession No.
1015945
Grant No.
2018-33610-28507
Cumulative Award Amt.
$96,835.00
Proposal No.
2018-00754
Multistate No.
(N/A)
Project Start Date
Aug 1, 2018
Project End Date
Mar 31, 2019
Grant Year
2018
Program Code
[8.5]- Food Science & Nutrition
Recipient Organization
SMART DIAGNOSTICS SYSTEMS LLC
7075 QUANTRILLS PASS
COLUMBIA,MO 65201
Performing Department
(N/A)
Non Technical Summary
Smart Diagnostics Systems (SDS) is creating its diagnostic system with a mission to protect consumers and companies from the dangerous threats and consequences that foodborne pathogens (ex. E.coli, Listeria, Salmonella, etc.) pose on a global basis. The outbreak of foodborne pathogens and foodborne illness is a significant and growing burden. The Centers for Disease Control estimates that each year nearly 48 million people get sick, nearly 130,000 are hospitalized and 3,000 die from foodborne pathogen illnesses, the biggest cause of illness in the United States annually. In addition to foodborne pathogens effects on the public, foodborne pathogen outbreaks and recalls costs food industry companies billions in losses every year. Due to the huge impact of foodborne pathogen outbreaks and foodborne illness, we are in great need of diagnostics that can rapidly, specifically, accurately, and cost efficiently detect foodborne pathogens throughout the food supply chain. This is why SDS is creating the PathoMeter, a state of the art diagnostic system (made up of a novel assay and optical detector) for the rapid detection of foodborne pathogens. In our field tests and pilot program SDS will be collecting data to validate the performance of the diagnostic system. Upon successful testing SDS will move forward to the commercialization of the diagnostic system. Commercialization of our diagnostic system will allow SDS to achieve its goal of preventing the outbreak of foodborne pathogens and the occurrence of foodborne illness. This pathogen detection breakthrough will revolutionize how we monitor, diagnose, and track the safety and quality of our food, ultimately protecting companies and saving lives from farm to fork.
Animal Health Component
30%
Research Effort Categories
Basic
20%
Applied
30%
Developmental
50%
Classification

Knowledge Area (KA)Subject of Investigation (SOI)Field of Science (FOS)Percent
71250102020100%
Goals / Objectives
The goal of Smart Diagnostics System's project is to create a detection system that significantly enhances the speed of foodborne E. coli O157:H7and Salmonella detection using customized selective antibodies for identification which will be synthesized on micro-magnetic platforms as optical platform. This will allow low volume and low concentration samples of pathogens to detect. The objectives of our project include:I. Design the Overall PathoMeter SystemBased on the targeted pollutant and its expected range of concentrations, SDS will define the major specifications of the system and design the overall system accordingly. We will model the system to optimize major design parameters. SDS will ensure that the design addresses USDA's most critical needs.II. Design and Develop System HardwareBased on the overall system design, SDS will design and develop major system components including optical sensor and control and readout electronics.III. Design and Develop System SoftwareSDS will design the control and data acquisition software for Phase I tests and demonstrations. The software will provide operation of the device and analysis of its measurements.IV. Design and Assemble Proof-of-Concept PrototypeSDS will assemble a breadboard proof-of-concept prototype system for Phase I feasibility tests. A laptop computer with a data acquisition board will be interfaced with the sensor chip through conditioning electronics.V. Demonstrate Feasibility of PathoMeter® SystemSDS will assemble a test setup and test the prototype system in the laboratory. The tests will use food with known pollutant concentrations through laboratory preparation. Field tests might be performed at farm sites. Upon successful testing, SDS will demonstrate the system's feasibility to USDA personnel.VI. Explore Commercial Potential and Product Viability to Prepare for Phase IISDS will explore the potential to transfer the PathoMeter® System biotechnology. Market research will identify the most promising applications of the PathoMeter® system.
Project Methods
The established Lab's standard protocol relies on the use of spherical micro-magnetic platform as the bone structure to assemble sandwich ELISA-based to capture and simultaneous detection of the target bacteria without the need for the elution step which aims at decreasing the steps and reduce time required in the testing process. Our protocol uses two sets of commercially available mouse or rabbit anti E. coli O157:H7 monoclonal total IgG antibodies. The capture antibody is unlabeled anti E. coli O157:H7 IgG antibody, which is pre-incubated with the protein A-coated magnetic beads in a concentration and time that ensures the saturation of the maximum binding capacity of the magnetic in accordance with the manufacturer's protocol. One the other hand, the detection antibody is an enzyme-linked (HRP-linked) anti E. coli O157:H7.When the target microbe is NOT present: The spherical micro-magnetic platform will be suspended in 1ml PBS cuvet. When 100μl of the prepared and enriched meat sample is added to the assay, the matrix will be incubated for 15 minutes to allow binding and reaction. The next step is to hold the micro-magnetic platforms with a magnet to remove the solution then wash the micro-magnetic platforms to remove any unbound substances. The next step is to add HRP-linked antibody and incubate for 15 minutes. The following step is to wash with PBS then add colorless HRP substrate. Since the target microbe is not present the color will not change.Microbe detection: As shown in the schematic representation of the testing steps, presence of the antigen (E. coli O157:H7) in the tested sample will result in capture of the target bacteria specifically by the spherical micro-magnetic platform and consequently binding of the HRP-linked detection antibody to the surface proteins of the viable bacteria. This will result in retaining of the HRP-linked antibody on the magnetic beads complex when the colorless HRP-substrate (TMB) is added, which in turn will produce blue/purple color in proportion to the amount of HRP present i.e., in proportion to the number of viable E. coli O157:H7 in the sampleThis biotechnology will help the end uses to test the safety of their products by selectively and accurately search for pathogens in the food product and get the results in around 8 hours. The evaluation will be conducted for each of the tasks performed.Task 1. Design the Overall PathoMeter®System Based on the targeted pollutant and its expected range of concentrations, SDS will define the major specifications of the system and design the overall system accordingly. We will model the system to optimize major design parameters.Task 2. Design and Develop System Hardware Based on the overall system design, SDS will design and develop major system components including optical sensor and control and readout electronics. The Biological Engineering at Lincoln University will assist with sensor development.Task 3. Design and Develop System Software SDS will design the control and data acquisition software for Phase I tests and demonstrations.The software will provide operation of the device and analysis of its measurements.Task 4. Design and Assemble Proof-of-Concept Prototype SDS will assemble a breadboard proof-of-concept prototype system for Phase I feasibility tests.A laptop computer with a data acquisition board will be interfaced with the sensor chip through conditioning electronics developed in Task 2.Task 5. Demonstrate Feasibility of PathoMeter® System SDS will assemble a test setup and test the prototype system in the laboratory. The tests will usefood with known pollutant concentrations through laboratory preparation. Field tests might be performed at farm sites. Upon successful testing, SDS will demonstrate the system's feasibility to USDA personnel.Task 6. Explore Commercial Potential and Product Viability SDS will explore the potential to transfer the PathoMeter® System biotechnology. Market research will identify the most promising applications of the PathoMeter® 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 08/01/18 to 03/31/19

Outputs
Target Audience:SDS's target customer is still protein and produce processing companies, but the company is evaluating other sectors of the food supply chain (e.g. manufacturing, distribution, and retail). The market research and interviews conducted to this point has allowed us to learn about food safety problems that exist within other sectors of the food supply chain, and their need for a solution. In addition to conducting market research for other potential applications in the food supply chain, SDS is also researching the potential and impact of our technology in countries outside of the United States. Food safety is a major problem in the United States, but it is a much larger problem outside of the United States. Because of this SDS is researching the impact our SBIR technology could have in developing and third world countries. Our immediate focus is still food processing companies in the United States, but if we can expand to other food industry segments and countries without altering or making significant changes to our current detection system it would make sense for us to expand our efforts. We will continue to conduct research on potential markets and technology applications for the duration of Phase 1. SDS has hosted workshops and laboratory instruction and is in the process of organizing more workshops for delivering science based knowledge. Because we have achieved successful feasibility testing, we hosted workshops and laboratory instruction to get feedback from industry stakeholders. We will continue to conduct workshops for the duration of Phase 1. We believe this will allow us to implement any modifications (from feedback) and solidify stakeholder relationships needed for Phase 2. 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?SDS has disseminated results and data, accumulated in Phase 1, through laboratory demonstrations and technology workshops. What do you plan to do during the next reporting period to accomplish the goals? SDS will continue to conduct market research to analyze how we can further develop our technology to address other critical needs in the food industry and USDA. SDS will explore further development of our hardware to address inefficiencies and problems currently experienced with conventional pathogen detection hardware. SDS will further validate the performance and success of the software while conducting testing for the duration of Phase 1. We will continue to test our prototype to ensure that the technology is ready to be implemented within industry for pilot testing in Phase 2. SDS will continue to collect testing data to prove and validate the performance and feasibility of our detection system. SDS will engage with food processing companies to review testing data accumulated in Phase 1 and determine the viability of SDS's detection system in its current state. If companies confirm that SDS's prototype is viable, we will attempt to secure pilots for Phase 2.

Impacts
What was accomplished under these goals? We have developed reagents (for strains of E.coli and Salmonella), hardware, and software. SDS has designed and developed the overall detection system. The designed detection system addresses the food industry's and USDA's most critical needs. Accomplishing this goal has allowed SDS to move forward in conducting feasibility testing and experiments to validate the performance (i.e. rapid detection time, accuracy, ease of use, etc.) of our detection system. SDS successfully developed the hardware using optical sensors and control and readout electronics. The hardware was designed to be portable, while achieving ultimate ease of use. The hardware has been used to conduct feasibility testing and collect the data generated from testing. SDS successfully developed software to conduct control and data acquisition for testing. The software has been used to operate the hardware and analyze the raw data generated from feasibility testing. SDS has designed and assembled the prototype of our detection system. The prototype was interfaced with a sensor chip and a laptop computer with data acquisition boards. The successful development of the prototype has allowed SDS to conduct experiments, testing and collect the data needed for to validate the feasibility and performance of our detection system. SDS has created an industrial testing simulation, within our laboratory, to conduct feasibility testing with our detection system. From industry interviews, we were able to replicate the operations, processes, and logistics of industrial testing, within our laboratory. Testing has been conducted with food samples subjected to known concentrations of contaminants (e.g. E.coli and Salmonella). SDS has accumulated successful data for feasibility testing to this point, and will continue to conduct testing for the duration of Phase 1 to further validate the potential for our technology to be implemented in industry. SDS has conducted 200+ customer discovery interviews in order to research and evaluate the best market applications for our detection system. From our research we have determined that the best, immediate, application of our detection system will be within food processing plants. The research and interviews conducted validated the commercial potential and viability of SDS's detection system. SDS evaluated each potential market by pain points/needs, resistance to change, total market opportunity, sales cycle, ability to scale, etc. and that's how we determined food processing companies would be our immediate target customer.

Publications


    Progress 08/01/18 to 03/31/19

    Outputs
    Target Audience:SDS's overall audience is companies in the food supply chain (i.e. Manufacturing, Processing, Distribution, and Retail). SDS's target audience is food companies that process ready-to-eat, fresh, and raw protein and produce products. SDS's detection system has proven to be scalable to various sizes of processing facilities. SDS's system can integrate and perform the same in a range of facilities, from a small independent processor to a large processor (e.g. Tyson, Cargill). This allows SDS to have large opportunity in our initial target audience. Within food processing companies we target lab technicians, food safety and quality assurance managers, and chief operations officers. SDS's work in industry has allowed us to identify that the listed individuals are the main influencers and decision makers for food safety products and services. During Phase I, SDS also evaluated the feasibility of expanding our detection system to the other sectors of the food supply chain. In our market studies and customer discovery interviews we evaluated our ability to integrate into other sectors of the food supply chain without significantly altering our current detection system. In our market development studies and research we concluded that our detection system could integrate into the other sectors of the food supply chain, but it could create a distraction to our immediate and target audience. From work conducted in Phase I we decided it would be best for SDS's success to focus on the U.S. processing market until we acquire a large market share, then expand to the other sectors of the food supply chain. In addition to our market work, we also evaluated the potential of expanding our technology internationally and into disadvantaged countries. The market need definitely existed internationally and we learned a high demand existed for rapid and accurate detection technologies. In the best interest of the company, we believe it will be best to first obtain a large share of the U.S. market before expanding internationally. Earlier expansion of our system would be a result of a joint venture or distribution partnership that wouldn't exhaust SDS's resources to expand products and services. SDS conducted many acts and processes that delivered science-based knowledge to industry stakeholders through technology demonstrations and webinars. Companies like Cargill, American Foods Group, Ecolab, and several independent processors attended demonstrations at SDS's facilities or webinars. The acts and processes allowed SDS to inform the industry on the development, features, advantages, functionality, and performance of our SBIR technology. The industry attendees provided feedback for adjustments we could incorporate to make the system more friendly and scalable in the food processing industry. The acts and processes led to SDS creating partnerships with many of the companies because of their interest in seeing the development and commercialization of SDS's technology. Changes/Problems: Nothing Reported What opportunities for training and professional development has the project provided?In Phase I SDS trained lab technicians to conduct benchmark testing using conventional methods (PCR) and SDS's SBIR technology. Many of the lab technicians were microbiologist familiar with molecular detection methods or lab techs that previously used other methods of detection. The training allowed SDS to evaluate ease of use and training time, while educating active testing personnel in the use and procedures affiliated with our detection system. The training allowed personnel to become more familiar with existing methods and were able to utilize the learned knowledge to improve certain aspects of SDS's system in Phase I. SDS conducted professional development for tech and business personnel. SDS's technology team attended conferences and webinars to learn more about testing regulations standards, future food industry legislation, modernization acts, and how to successfully accredit SDS's system for wide scale acceptance across the industry. The business team participated in accelerators and incubators like VentureWell and Techstars to advance SDS's knowledge of our market, commercialization plans, manufacturing strategy, maximizing margins, and maximizing potential to rapidly scale. How have the results been disseminated to communities of interest?SDS has disseminated our results from Phase I through webinars, technology demonstrations, and presentations. The results were disseminated to the science community through webinars, food processors and industry stakeholders through technology demonstrations, and to the public through presentations. 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 Phase I SDS developed reagents (for strains of E.coli and Salmonella), hardware, and software. SDS has designed and developed the overall detection system. The designed detection system addresses the food industries and USDA's most critical needs. Accomplishing this goal has allowed SDS to move forward and conduct testing and experiments to validate the feasibility and performance (i.e. rapid detection time, accuracy, ease of use, etc.) of our detection system. SDS successfully developed the hardware using optical sensors and control and readout electronics. The hardware was designed to be portable, while achieving ultimate ease of use. SDS achieved advances in automation and higher sample throughput. The automation will allow SDS's testing process and steps to be autonomously conducted within the instrument. SDS also developed a carousel within our instrument that will allow us to test between 5-30 samples in a single analysis. The hardware has been used to conduct feasibility testing and collect the data generated from testing. SDS successfully developed software to conduct control and data acquisition for testing. The software has been used to operate the hardware and analyze the raw data generated from feasibility testing. SDS also developed interfaces that allows industry partners to access testing data to provide feedback and analysis of testing results, testing protocols, and system performance. SDS has designed and assembled the prototype of our detection system. The prototype was interfaced with a sensor chip and a laptop computer with data acquisition boards. The successful development of the prototype has allowed SDS to conduct experiments, testing and collect the data needed for to validate the feasibility and performance of our detection system. SDS has created an industrial testing simulation, within our laboratory, to conduct feasibility testing with our detection system. From industry interviews, we were able to replicate the operations, processes, and logistics of industrial testing, within our laboratory. Testing has been conducted with food samples subjected to known concentrations of contaminants (e.g. E.coli and Salmonella). SDS has also tested "blind" and real world samples to conduct testing. SDS's testing results were validated through plating the samples. All SDS's testing in Phase I was confirmed accurate by plating confirmation. Through Phase I the system provided results that were 100% accurate and experienced no false positive or false negative results. SDS has conducted 215+ customer discovery interviews in order to research and evaluate the best market applications for our detection system. From our research we have determined that the best, immediate, application of our detection system will be within food processing plants. The research and interviews conducted validated the commercial potential and viability of SDS's detection system. SDS evaluated each potential market by pain points/needs, resistance to change, total market opportunity, sales cycle, ability to scale, etc. and that's how we determined food-processing companies would be our immediate target customers. SDS established numerous food industry partnerships that will aid SDS in successfully implementing our SBIR technology into the food processing industry.

    Publications