SINGLE-TUBE CAPTURE, CONCENTRATION, AND GENOMIC EXTRACTION OF FOODBORNE VIRUSES USING MAGNETIC IONIC LIQUIDS

GOALSTheoverall goalof this work is to evaluate the potential of an emerging technology, namelydispersive droplet magnetic ionic liquid (MIL) extraction,to perform rapid, portable concentration and genomic extraction of non-enveloped viruses from contaminated food matrices, thus lowering current barriers to routine pathogenic virus testing in food and environmental samples. The specific goals are as follows:1: Develop capture and concentration protocols for non-enveloped virus and viral ssRNA using MILs that do not require electrical equipment.Current methods rely on vortex mixing for dispersion of MILs in suspension. Here we will explore using a Pasteur pipette to disperse MIL droplets for target capture and elution, eliminating the reliance on electrical equipment. We will also evaluate factors impacting the separation of MILs from suspension by an external magnetic field, since this aspect eliminates the need for centrifugation.2: Evaluate the potential of MILs as binding substrate for viral ssRNA extraction.In this aim we will explore the possibility of single-tube virus capture and ssRNA concentration. This will involve development of shelf-stable reagents for capsid lysis, wash, and elution of target ssRNA that do not damage the MIL structure or disrupt target binding.3: Evaluate performance of a MIL-based target capture and concentration protocol in the presence of complex food matrices.In this aim we will explore the impact of complex food matrices on virus recovery by MIL extraction. Specifically, we will be testing artificially contaminated romaine lettuce, frozen raspberries, and oysters, since these foods have each been frequently associated with HuNoV outbreaks.?