Project summary: The supplement application is in response to the RADx-rad announcement on the opportunity of additional resources and funding support to facilitate the pathway to commercialization. As part of this supplement we will pursue specific milestones to de-risk the Rolosense technology and further move it toward commercialization. A major goal will be to introduce multiplexing capabilities to simultaneously detect multiple viral targets from the same sample. Encoded Rolosense particles with unique virus-binding ligands will be employed to achieve this goal. Multiplexing to detect multiple viral targets will help distinguish the capabilities of Rolosense from that of the current state-of-the-art. Another goal is to increase the speed and robustness of the assay to facilitate direct sensing from breath condensate samples. This second goal will be pursued by further developing fuel-free Rolosense that eliminates the need for RNA and RNaseH enzyme in the assay. We see this as a critical step to reduce the cost of the assay and to increase rigor and reproducibility to achieve the commercialization goals without concern for RNaseA found in most biological samples.