AqPathDx: Simple and Easy to Use Diagnostic kit for Rapid Detection of Catfish and Tilapia Pathogens

Varizymes proposed AqPathDx kit enables rapid pondside detection of the most common catfish and tilapia pathogens in two separate panels in less than 30 minutes using a novel multiplex molecular diagnostic assay. This capability provides a critical unmet need for aquaculture and meets USDA Strategic Goal 2 in ensuring America's agricultural system is equitable resilient and prosperous as well as USDA Science and Research Strategy priorities 1 and 4. The AqPathDx kit is based on multiplex loop-mediated isothermal amplification (mLAMP) method which allows real-time detection of up to 4 targets in a single reaction. This assay will be performed on a benchtop isothermal instrument and the results will be displayed on the screen as positive negative or non-conclusive. AqPathDx kit performance will be compared with that of the reference method. A simple and rapid sample preparation method will also be developed and complete reaction formulation will be lyophilized for ambient storage. The analytical performance of the AqPathDx kit will be evaluated by testing a cohort of 300 positive and 200 negative samples. Successful completion of Phase II will lead to: • Development of a diagnostic kit (AqPathDx) for the detection of multiple pathogens at pondside. • Total assay time of 30 min including sample preparation. • Simple and easy-to-use sample preparation method. • Diagnostic sensitivity and specificity comparable to reference methods. • Lyophilized reagents for storage at ambient temperature. • Platform technology for rapid detection of other pathogens.