Development of HyBeacon® probes for the forensic detection of Panthera, rhinoceros, and pangolin species.

The Illegal Wildlife Trade (IWT) represents a multi-billion dollar black-market industry whereby wild species are illegally taken from their natural environment and sold. A common question asked by wildlife forensic scientists pertains to species and/or genus identity, which currently requires multi-step processing. Our work details the development of three HyBeacon® probes, used for the presumptive detection of rhinoceros, pangolin and key target species in the Panthera genus. The approach can be performed in a single tube using melt curve analysis and provide rapid assessment of sample identity. Using synthetic DNA of representative species, early data suggest the approach is sensitive enough to achieve species identification with <10 cells. Future development and assay validation can allow the rapid screening of multiple seized items before confirmatory DNA sequencing.