Rapid environmental DNA detection with isothermal amplification methods: a comparison with the real-time quantitative PCR in the natterjack toad (Epidalea calamita).

RPA matches qPCR sensitivity for detecting Epidalea calamita eDNA from aquatic filtrations, while LAMP is ten times less sensitive.

• Why it matters: PCR-based eDNA detection is often slow and lab-dependent, leaving a gap for rapid, field-deployable alternatives with comparable sensitivity and specificity.
• What they did: The study designed LAMP and RPA primers from the mitochondrial genome, tested sensitivity and specificity on DNA extracts and tissues using fluorescence readouts, then validated all assays on pond eDNA where field scientists observed the species.
• The result: In eDNA from surveyed ponds, all assays amplified E. calamita DNA when the species was observed, and RPA’s sensitivity was equivalent to qPCR, enabling robust isothermal eDNA workflows under constrained time and lab access.

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