The aim of DxHub is to develop a strong ecosystem that mutually reinforces actors from two widening countries to consolidate individual efforts and allow for rapid and on the spot accurate diagnosis of infectious diseases within One-Health. The ecosystems in Crete, GR and the Oeiras region, PT, have already demonstrated a rich portfolio of activities to tackle the problem of efficient diagnosis outside a centralized lab. The DxHub will bring the two isolated regions together and, through a common R&I plan, accelerate innovations in diagnostics into impactful solutions by embracing the quadruple helix of main innovation players. Cutting-edge R&Is will be promoted based on joint research, innovation and investment plans. Pilot projects will be used to interlink academia with business providing solutions addressing the health, veterinary and agri/food sectors, all in line with both regions’ strategic priorities. In parallel, the efficient communication, dissemination and exploitation of the DxHub outcomes to the broad ecosystem will ensure high societal impact as a result of the co-development and acceptance of the new innovations by all actors of the quadruple helix. We expect that by building strong links between all payers of DxHub will foster knowledge transfer leading to the creation of new jobs and of attractive and sustainable place-based innovation ecosystems.