Periodic Reporting for period 1 - DART Diagnostics (Saving Time, Money and Lives Through Rapid Plug & Play Salmonella Diagnostics) Reporting period: 2018-09-01 to 2019-02-28 Summary of the context and overall objectives of the project With multiple outbreaks of foodborne illnesses and massive product recalls witnessed during the last years, the current state of the art food testing technologies still require 3-5 days to detect major food poisoning agents like Salmonella. However, supply chain pushes food manufacturers to send out products to retailers well before potential pathogens can get detected. DART is a fast detection kit of the Salmonella bacteria for food safety applications, based on a unique amplification reagent originally developed by us and enabled by the latest nano-medical advances. The DART Diagnostics project has as objective to improve immunoassays sensitivity, and allow for faster and more accurate diagnostics. Early detection of Salmonella allows the technology to be employed in the food industry, which has lower entrance barriers (than the clinical diagnostics), namely in terms of the certifications and validations needed, implying lower regulatory costs. Work performed from the beginning of the project to the end of the period covered by the report and main results achieved so far During the Phase 1 project, the work that has been completed covered by the report is summarized in the following bullets:• We executed a comprehensive food safety market assessment of hypothesis and assumptions combined with an analysis of environmental forces, market trends, competition and risks, opportunities and comprising our resources and constraints. • We have identified our end-users and potential clients in the key markets we are currently targeting, which are essential to analyse the current trends in the food safety market.• We performed a comparative in-depth analysis of the best countries to target and what would be the best commercialisation strategy as well as drafting a concise plan on overcoming any market entry obstacles we might face.• We secured our Freedom to Operate in our key markets through extensive patent research. • We have drafted a realistic and attainable work plan for the further development of our kits and technology to ensure that all the objectives are met, focusing our attention on specific technical and commercial obstacles that could present limitations to our market introduction. Progress beyond the state of the art and expected potential impact (including the socio-economic impact and the wider societal implications of the project so far) The focus of our work has been to amplify the signal from classical immunodiagnostic assays, by using nanomaterials first designed at the Instituto de Medicina Molecular, Faculdade de Medicina, Universidade de Lisboa, Lisbon, Portugal and fully developed at DART Diagnostics, Tec Labs – Campus of the Faculty of Sciences of the University of Lisbon (FCUL), Lisbon, Portugal. The nanomaterials developed and used are intrinsically biocompatible, stable in physiological conditions and the basis of a promising new technology, which was employed in immunoassays, to achieve high sensitivity and fast results. Several experimental conditions were tested, seeking to find the most suitable modifications to the detection protocol, in order to allow it to be used for diagnostics applications in the future. This information was then used to detect a target protein, the glial fibrillary acidic protein (GFAP) as well as Salmonella spp. bacteria. The technology can still be improved, both in terms of the time length of the protocol and in terms of sensitivity, to detect even lower quantities of the target protein. Such improvements will allow it to be employed in foodborne pathogen detection and in clinical diagnostics in the future. GFAP presence in the blood can be used for the earlier detection and classification of stroke, being thus clinically relevant as a disease biomarker.