Project will address an urgent need for field-based detection tools for harmful invasive microbes and work with end- users to develop, validate, and deploy three practical and reliable LAMP assays.
Rapidly responding to the invasion of harmful microbes can be limited by insensitive and labor-intensive methods or require highly technical laboratory capacity. To address these limitations, we will develop a sensitive and field-deployable molecular tool for detecting DNA and RNA in the environment using Loop-mediated Isothermal Amplification (LAMP). In consultation with end-users, we have identified three high-priority microbes of interest for LAMP assay development: viral hemorrhagic septicemia virus (VHSV), largemouth bass virus (LMBV), and Didymosphenia geminata (Didymo). This project aims to validate and field test new assays for each microbe and build management capacity to support ongoing surveillance efforts. We will first optimize (VHSV and LMBV) and develop (Didymo) lyophilized LAMP assays for the three high-priority microbes. Each assay will be validated under laboratory conditions to determine specificity and sensitivity. Then, we will deploy the validated assays in the field by testing known-positive locations in Minnesota (LMBV and Didymo) or virus-exposed fish in the laboratory (VHSV) to evaluate detection limits compared qPCR. Field testing will done in partnership with management end users to evaluate practicality/usability of the LAMP assays and build capacity using standardized SOPs.