Location: Produce Safety and Microbiology Research
Project Number: 2030-32000-011-005-I
Project Type: Interagency Reimbursable Agreement
Start Date: Oct 1, 2022
End Date: Sep 30, 2027
Objective:
This project aims to improve animal surveillance diagnostic capability through the development of a direct, rapid, sensitive SARS-CoV-2 detection tool that can be readily utilized in the field and fill the gap by offering in-field point-of-care (POC) COVID-19 diagnostics tests for animals. The objective of this project is to develop an aptamer-based lateral flow assay (aLFA), which is a pen-side diagnostic tool that can be used and deployed as an early pre-clinical detection test through animal swabs and environmental sampling. Specifically, this proposal addresses and responds to the immediate threat presented by SARS-CoV-2 by 1) generating highly specific and stable aptamer sequences that can bind to SARS-CoV-2 variants and integrating onto a LFA system for the direct detection, 2) optimizing aLFA conditions to establish a robust diagnostic system which can efficiently respond to new variants as they occur, and 3) applying the newly developed aLFA diagnostic technology on animal and environmental samples for validation purposes, routine testing, and surveillance and monitoring programs.
Approach:
ARS scientists will (Stage I) generate in-house aptamers to serve as capture and detection elements by conducting SELEX (systematic evolution of ligands by exponential enrichment) and using viral recombinant proteins of SARS-CoV-2 variants of concerns (VOCs) and DNA library. Stage II will be focused on constructing and optimizing the aLFA system which is consisted of the following major components: sample application pad, conjugate pad, nitrocellulose membrane, absorption pad, one-hole cassette, and the highly-specific aptamers generated from the previous stage. Stage III of the project will be on the application of the aLFA system to evaluate its stability and determine its limit of detection on animal and environmental samples.
