An Aptamer-Quantum Dot-Based Lateral Flow Test Strip for Cyclospora Detection

Phase II Technical Objectives (TOs): TO-1 Identify extant DNA aptamer sequences from Phase I aptamer pools or develop new DNA aptamers that clearly bind the outer cell wall surface of unsporulated Cyclospora cayetanensis oocysts, since the current aptamers appear to stain spores inside sporulated oocysts, but perhaps also the cell wall surface. Some current aptamers may also detect unsporulated oocysts upon further screening, but if not, Nanohmics will develop whole oocyst aptamers to bind the outer cell wall. Note: the paucity of available oocysts from the CDC in Phase I, limited Nanaohmics' ability to screen aptamers for oocyst surface staining by fluorescence microscopy.TO-2 Fully develop lateral flow (LF) aptamer-quantum dot (Qdot) test strips and assess their limits of detection (LODs) for sporulated and unsporulated Cyclospora cayetanensis oocysts.TO-3 Develop a large-scale aptamer-fluorescence filter test and determine its LOD for Cyclospora oocysts from 10-50 or more liters of water.TO-4 Compare, downselect and build a prototype LF test strip fluorescence reader or automated fluorescence microscope for large volume filters.TO-5 Interface with agricultural business leaders (e.g., Conagra, Xgenex, etc.) and LARTA to develop a licensing agreement or other business/commercialization strategy.TO-6 Submit quarterly and/or annual and final technical reports as required or desired by USDA NIFA.