Western Australia used a systematic triage approach to verify eDNA detections of the invasive colonial ascidian, Didemnum vexillum, resulting in the first confirmed detection in Australia. As the popularity of eDNA technologies evolves, it is vital to have a robust process in place to verify eDNA-based detections prior to initiating large-scale responses. eDNA detection does not necessarily confirm the presence of a species, and the reverse is also true, as it comes with numerous caveats. We draw on the recent example of D. vexillum detection in Western Australia to outline the significance of incorporating a triage approach. We also highlight the importance of establishing a fit-for-purpose, in-house reference collection unique to the jurisdiction, to navigate the complexities of taxonomic identification and molecular false positive challenges with species such as D. vexillum. The approach taken through this emergency pest response underscores the critical need for systematic verification processes and comprehensive reference libraries to ensure accurate, timely and reliable eDNA detections, ultimately aiding in effective biosecurity management.