Development of quantitative PCR primers and probes for environmental DNA detection of amphibians in Ontario

DNA from environmental samples (eDNA) is increasingly being used to detect and monitor elusive species. We developed species-specific eDNA primers and probes for qPCR detection of 24 amphibian species native to Ontario, Canada. Cross-species testing confirmed their high specificity and low cross-species amplification, as well as their ability to detect DNA from target species at low concentrations. These detection tools should prove useful for monitoring at-risk amphibian species throughout their native ranges.