These Environmental DNA test validation guidelines provide harmonised quality control and minimum standard considerations for developing or validating eDNA/eRNA assays for the purpose of single-species or multi-species detection. This document is a comprehensive guide for the development and use of eDNA/eRNA tests, recommended and curated by eDNA experts, stakeholders and end users in Australia and New Zealand. The guidelines are designed to support a consistent and best-practice approach to eDNA/ eRNA testing to help detect species of interest. This approach ensures that surveillance and resource managers are provided with robust scientific evidence to support decision making.