A novel approach to combatting proboscidean ivory trafficking using a multiplex High-Resolution Melt (M-HRM) assay.

To support efforts in prosecuting wildlife crimes, we developed and validated a multiplex High-Resolution Melt (M-HRM) assay for the identification of proboscidean taxa commonly required to be identified or excluded in ivory seizures and forensic casework: Asian elephant (Elephas maximus), African elephant (Loxodonta spp.), mammoth (Mammuthus spp.), and mastodon (Mammut spp.). Five hundred and fifty (550) blood, tissue, and ivory samples from individuals of these 4 proboscidean taxa were used to develop and validate the 2 proboscidean-specific mitochondrial sites targeted by this assay. The 28-basepair (bp) 16S ribosomal RNA (rRNA) and 54-bp cytochrome b (Cytb) gene segments yield a combination of melt peaks that create composite melt profiles unique to each of the 4 proboscidean taxa. Wildlife forensic laboratories can use this sensitive, rapid, and cost-effective assay to assist efforts to combat the unlawful commercialization of proboscidean ivory and to stop the poaching crisis leading to the decline of these ivory-bearing species in the wild.