Potyviruses, members of the genus Potyvirus, collectively infect almost every major crop throughout the world, and in many crops a potyvirus may be the most economically important viral pathogen. Efficient management of viral diseases requires detection and identification tools and protocols for quick, reliable and accurate identification of the pathogen. Several years ago, the USDA-ARS-USNA Beltsville Lab developed a hybridoma cell line that secretes a broad-spectrum reacting PTY-1 monoclonal antibody (McAb), which detects almost all known aphid-transmitted potyviruses, was patented and licensed to Agdia, Inc., and forms the basis of Agdia’s “POTY Group Test” reagents, ELISA kits, and ImmunoStrip Test. CANARY^TM (Cellular Analysis and Notification of Antigen Risks and Yields) is a fast, sensitive, and easy-to-use biosensor-based technology that has been successfully utilized for the detection of bacteria, viruses, and toxins. Several PTY-1 hybridoma-derived CANARY biosensor clones were created at MIT-LL, initially tested in the APHIS-CPHST Beltsville Lab, and later evaluated collaboratively in the PathSensors and USDA-ARS labs, for assay feasibility, bead capture efficacy, specificity and sensitivity of the reaction to detect target potyviruses. The optimized POTY biosensor can detect diverse purified potyviruses down to less than 5 ng/ml and can detect potyviruses in naturally infected sources, such as various isolates of potato virus Y in potato leaves and tubers.