Foodborne illnesses devastate the gastrointestinal microflora of millions of Americans each year, often resultingin dysbiosis and increase risk of other diseases, namely irritable bowel syndrome, kidney failure, pancreatitisand diabetes. Furthermore, diabetics are at increased risk of developing a foodborne illness, and they often takelonger to recover. Each year, 1 in 6 people in the U.S. are sickened by foodborne pathogens. In 2014, therewere 19,542 laboratory confirmed infections, 4,445 hospitalizations and 71 deaths attributed to the 9 majorpathogens transmitted commonly through food. Listeria is the one of the most serious offenders, with a 96%hospitalization rate and a 12.9% mortality rate. The impact on healthcare, business and industry is no lesspronounced, resulting in annual costs of $14.1 to $16.3 billion, including direct medical costs and value of timelost to illness. Moreover, the cost of keeping food safe from Listeria contamination was estimated to rangebetween $2.4-$2.6 billion. Contaminated food is the cause of these illnesses, having entered the public supplydue to a lack of detection at a manufacturing/packaging plant, distribution warehouse or retail location. Currenttests are inadequate because they require a prolonged enrichment step to reduce the likelihood of falsenegatives and false positives. This enrichment causes tremendous delays in providing actionable results to thefood processor, who are left with one of two choices: Store the products until the test result comes back, resultingin older, lower-quality food; or shipping the food before receiving test results, putting the consuming public atincreased risk. Sample6 is developing the world?s first enrichment-free foodborne pathogen detection system.DETECT/L is a rapid screening assay that detects a single Listeria spp. cell on environmental surfaces in lessthan one work shift (<7h). Sample6?s proprietary Bioillumination Platform enables this expeditious turnaround,enabling food manufacturers and packagers to quickly and accurately detect and remediate contaminatedenvironmental surfaces, and solely retain product that is at risk of contamination. However, food can becomecontaminated internally during production, shipping, or in wholesale and retail environments downstream in thesupply chain. It is of vital importance to develop a second testing modality that assays the foodstuff as rapidlyand sensitively as DETECT/L does for environmental surfaces. To this end, the proposed Phase II study willcontinue the work begun in Phase I to adapt the DETECT/L assay for finished product testing. First, theresearchers will expand the number of Listeria strains to 10. Next, assay development will be completed for thesix foods begun in Phase I, plus 30 more foods. Finally, Sample6 will develop high-throughput methods so thatfood manufacturers and packagers can test multiple food samples at a time for efficiency.