IMPROVING THE LIMIT OF NOROVIRUS DETECTION IN FOOD MATRICES USING ENGINEERED YEASTS TO CONCENTRATE AND PURIFY VIRAL PARTICLES

In this proposal, we aim to genetically engineer yeast to display specific virus-binding single-domain antibodies (called nanobodies) with high capture efficiency on the cell surface, facilitating the concentration and purification of human noroviruses in food matrices. The binding nanobodies on yeast surfaces can be programmed to achieve a relatively high capture efficiency and/or serve as anchors to capture different norovirus genogroups. In this study, food-grade yeasts (Saccharomyces cerevisiae,EBY100) will be engineered to display specific virus-binding nanobodies that can specifically bind to human noroviruses. After binding, human noroviruses in complex food samples can be easily concentrated and purified by filtration or centrifugation of the much more massive yeast cells. RT-qPCR will then be used to estimate capture efficiency and detect human norovirus. Herein, thehypothesis of this workis that engineered yeasts can serve as the next generation of foodborne contaminant-specific substrates for the easy concentration and rapid detection of human noroviruses in food samples. This study would not only be a revolutionary novel strategy to detect foodborne viruses but will also be amongthe most efficient and cost-effectivetechniques for norovirus detection.