Global biodiversity targets require us to identify species at risk of extinction and quantify status and trends of biodiversity. The Red List Index (RLI) tracks trends in the conservation status of entire species groups over time by monitoring changes in categories assigned to species. Here, we calculate this index for the world’s fishes in 2010, using a sampled approach to the RLI based on a randomly selected sample of 1,500 species, and also present RLI splits for freshwater and marine systems separately. We further compare specific traits of a worldwide fish list to our sample to assess its representativeness. Overall, 15.1% of species in the sample were estimated to be threatened with extinction, resulting in an sampled RLI of 0.914 for all species, 0.972 in marine and 0.860 in freshwater ecosystems. Our sample showed fishing as the principal threat for marine species, and pollution by agricultural and foresty effluents for freshwater fishes. The sampled list provides a robust representation for tracking trends in the conservation status of the world’s fishes, including disaggregated sampled indices for marine and freshwater fish. Reassessment and backcasting of this index is urgent to check the achievement of the commitments proposed in global biodiversity targets.