Project Summary/Abstract Each year over one million people become ill due to foodborne Salmonella infections, this accounts for a total of 11% of all foodborne illness reported in the United States1. Salmonella is the leading cause for foodborne illness hospitalizations (23,000 cases; 35%) and deaths (450 cases; 28%). Salmonella has an enormous impact on the economy, causing $3.7 billion in costs in the United States each year; 90% of this burden ($3.3 million) is due to deaths, 8% ($294 million) due to hospitalizations and 2% due to non-hospitalized cases. The primary mode of Salmonella infection is through food, mainly raw or undercooked meat, raw eggs and unwashed vegetables. These illnesses are often tied to outbreaks that result from contaminated food that was not detected at a manufacturing or packaging plant. Current detection methods for Salmonella provide slow time to actionable results, making it difficult to take necessary steps in controlling the pathogen. Sample6 has developed a new technology, based on the proprietary Bioillumination PlatformTM, which utilizes a novel synthetic engineering approach to leverage the natural specificity of bacteriophage to detect the presence of specific bacterial species. Sample6 DETECT/S system contains engineered bacteriophages, which can identify Salmonella spp. to induce production of luminescent enzymes, upon infection of the target cells, effectively illuminating Salmonella. This signal can be measured with the Sample6 Luminometer and instantiates the world's first enrichment-free pathogen diagnostic system. Sample6 has previously established feasibility that the DETECT/S assay can be adapted for finished product testing to detect Salmonella directly on food products that are prone to contamination. For this Direct-to- Phase II project, we propose to complete the development of our technology to detect Salmonella on additional foods to achieve validation as laid out in AOAC guidelines. We will expand the testing to include 400 Salmonella isolates belonging to clinically relevant serotypes, and complete assay development for 20 food matrices representing all commercially relevant categories, including low-moisture foods. Finally, we will develop non-replicating phage technology for our BioIllumination Platform to further increase our reach in the market, both domestically and internationally.