Location: Produce Safety and Microbiology Research
Project Number: 2030-32000-011-008-N
Project Type: Non-Funded Cooperative Agreement
Start Date: Jan 1, 2023
End Date: Dec 31, 2025
The objectives of this cooperative research project are to (1) develop a lateral flow assay (LFA) for the simultaneous detection of pathogens, (2) develop an App for measuring and collecting the results of the developed LFA, and (3) investigate pathogens by applying the LFA kit and App which will then be further utilized to develop a risk assessment. Patent application and publications are the immediate potential outcomes of this collaboration which will provide ARS a stronger leverage to engage with the Bay Area biotech industries for potential technology transfer (TT) and commercialization of LFA system and other future assay developments. The LFA/App-based assay in a portable device format will improve the current detection methods of pathogens by providing the U.S. farmers an effective and reliable on-site/in-field screening wireless technology that can be employed in field. Several rapid screening solutions based on antibody or polymerase chain reaction (PCR) technologies are currently available on the market; these existing solutions, however are not cost-effective (ie PCR machine, monoclonal antibodies) and still experience cross-reactivity (polyclonal antibodies) and instability. LFA/App-based assay would allow these commodities to be readily screened on-site for the presence of pathogens, thus immediate results are generated that would save time (important in the event of outbreaks), cost and energy. This option is also expected to generate accurate qualitative and quantitative results, thus contamination levels are better estimated for a more precise assessment. Since the planned LFA/App-based assay will integrate aptamers instead of immune-based and PCR platforms, its cost is greatly reduced; procedures are more direct and less cumbersome, and results are easily interpreted using the App. It is also anticipated that a wider group of users from various fields, regulatory agency and research institution can readily adopt and apply it for routine testing and/or inspection in the USA. This improvement of methodology has a direct positive impact on the screening process of pathogens by reducing testing time and cost.
To achieve the objectives of this research project, the Cooperator will perform the laboratory experiments: (1) synthesize and produce aptamer-tagged gold nanoparticles (2) develop LFA kit (3) artificially inoculate pathogens. The Cooperator has the expertise in terms of developing both the aptamer magnetic nanoparticle-LFA kit system and its App which ARS does not have. The LFA device will be comprised of sampling and conjugate pads, nitrocellouse membrane, and absorbent pad. The capillary action will allow the migration and specific binding of samples and reagents which include the aptamer-tagged gold nanoparticles solution produced in the Cooperator’s laboratory. Therefore, generating preliminary data for proof of concept as well as optimization trials are reasonable to be conducted in the Cooperator’s laboratory. Both ARS scientists and the Cooperator will jointly (1) design the experiments, (2) analyze data, (3) develop the App specifically for the LFA kit system to collect, process, and interpret the data/signal generated, and (4) establish and perform risk assessment using the newly-developed LFA/App-based kit system. By working with the University/Cooperator, ARS will be able to develop its capability in the App development for LFA and explore other potential LFA-compatible detection elements such as bacteriophages, etc. which can be used as alternatives to the commonly utilized immune and molecular-based platforms.