Collaboration plays a pivotal role in our National Biosecurity System. Fostering the growth of networks and trusted relationships both domestically and internationally has improved our preparedness for both exotic and endemic pathogens. A collaborative approach was employed for Hort Innovation project MT17006, focused on increasing Australia’s preparedness for our number one High Priority Plant Pathogen. The journey started from humble beginnings with Australian State diagnosticians collaborating with New Zealand MPI on a project application for improving diagnostics for Xylella fastidiosa. This blossomed into sharing of DNA from exotic cultures, co-development of new diagnostics with validation. We witnessed firsthand the real-life devastation of Xylella fastidiosa through the development of overseas collaborations. Knowledge was shared, we were taught how to identify symptomatology in the field, and process samples for both the isolation and molecular detection of this exotic pathogen. Subsequently, collaboration saw the development of innovative diagnostics tools capable of identifying both the presence of Xylella at the genus level and the identification of the subspecies and sequence type with MinION sequencing. These and other methods were delivered as a new National Diagnostic Protocol (NDP) for Xylella spp., plus delivery of a workshop at the Australasian Plant Pathology Society Conference for diagnosticians and surveillance officers. All of this developed knowledge is being distributed and used in development of surveillance protocols for this pathogen and potential hosts. This project has seen techniques learned from overseas colleagues be applied to current endemic pathogens that are having an impact on Australian industries now.