The Murray–Darling Basin Authority, supported by state governments is responsible for reporting on the condition of the Basin, including the diversity and distribution of native fish. In 2021 MDBA launched the Basin Condition Monitoring Program, a $7.5 million 4-year monitoring program. Environmental DNA was identified as an innovative technology worth exploring amongst current fish monitoring techniques. A suite of eDNA projects is underway, including: i) review of eDNA projects that have occurred in the Basin; ii) eDNA for citizen science biodiversity surveys; iii) potential for species-specific assays to identify threatened species presence; iv) the efficacy and cost of species-specific vs metabarcoding for threatened fish species; and v) comparing species detection by eDNA vs traditional methodologies. A community of practice was established, to bring together researchers, public and private labs and government agencies using eDNA across the Basin. Twenty-seven eDNA projects within the Basin have been published since 2016, with 21 focused on fish eDNA. Citizen science sampling occurred at over 300 sites, detecting 41 different fish species, including 6 threatened species. Sixty-three sites were sampled for threatened fish across inland NSW and Queensland, with both species specific and metabarcoding analysis underway. eDNA samples have been collected at 110 electrofishing sites, with preliminary eDNA results identifying over 40 fish species. Over 150 sites are being sampled across the Ovens-Murray-Kiewa Rivers in spring/summer 2024 in partnership with DEECA, Latrobe University and NESP. In Addition, over 700 sites sampled in the Victorian MDB and 384 in the NSW MDB contribute to Basin Plan evaluation. This presentation examines early results of eDNA metabarcoding surveys for fish in the MDB and assesses the technique’s utility as a tool for monitoring fish biodiversity and species distribution in an area of over a million square kilometres.