Surveys of environmental DNA (eDNA) have become an important and multifaceted tool for monitoring and identifying distributions and occupancy of aquatic species. This tool is attractive because it is powerful, easy to apply, and provides an alternative to traditional field survey methods. However, validating eDNA survey methods against traditional field survey methods is warranted prior to their application. We used eDNA and electrofishing to survey 10 sites in 3 tributaries of the Chehalis River, Washington, to infer distribution and occupancy of Entosphenus tridentatus and Lampetra spp. Both methods produced similar detection rates for E. tridentatus, and Lampetra spp. were detected at slightly greater frequency with eDNA in the Black River and Skookumchuck River. Within each of the three tributaries, eDNA concentration was negatively related to sample distance from the Chehalis River mainstem for E. tridentatus but not for Lampetra spp., which indicates E. tridentatus and Lampetra spp. may be distributed differently within tributaries. Application of lamprey eDNA data to a multiscale occupancy model indicated high probability of detecting eDNA in water samples and quantitative PCR (qPCR) assays. Broad distribution and high detection of E. tridentatus and Lampetra spp. suggest robust populations inhabit the Chehalis River basin. Our findings suggest eDNA surveys may be comparable to electrofishing for informing lamprey occupancy and distributions. Such sampling is efficient and cost‐effective and we anticipate that eDNA surveys will become a valuable tool in addressing key research and monitoring needs for conservation and restoration of lampreys in general.