salamandrivorans in Amphibian Samples

Chytridiomycosis is a lethal fungal disease contributing to declines and extinctions of amphibian species worldwide. The cur-rently used molecular screening tests for chytridiomycosis fail to detect the recently described species Batrachochytrium sala-mandrivorans. In this study, we present a duplex real-time PCR that allows the simultaneous detection of B. salamandrivorans and Batrachochytrium dendrobatidis. With B. dendrobatidis- and B. salamandrivorans-specific primers and probes, detection of the two pathogens in amphibian samples is possible, with a detection limit of 0.1 genomic equivalent of zoospores of both patho-gens per PCR. The developed real-time PCR shows high degrees of specificity and sensitivity, high linear correlations (r2> 0.995), and high amplification efficiencies (>94%) for B. dendrobatidis and B. salamandrivorans. In conclusion, the described duplex real-time PCR can be used to detect DNA of B. dendrobatidis and B. salamandrivorans with highly reproducible and reli-able results. Chytridiomycosis causes worldwide declines and extinctions ofamphibian populations and is one of the most important in-fectious diseases in amphibians (1–3). Batrachochytrium dendro-batidis was the sole Chytridiomycetes taxon known to infect verte-brate hosts and to be able to cause this devastating disease (4) until a second chytrid species was isolated from a mortality event that drove the Dutch fire salamander (Salamandra salamandra