A tale of the traded cat: development of a rapid real-time PCR diagnostic test to distinguish between lion and tiger bone

A rapid allelic discrimination real-time polymerase chain reaction (qPCR) assay has been developed for the confirmation of lion ( Panthera leo ) or tiger ( P. tigris ) DNA. The method includes a low cost allele specific PCR assay designed for each SNP variant where the genotype is determined by incorporation and subsequent fluorescence of FAM or HEX following PCR. The described method was able to accurately and rapidly distinguish between the two species, and all three mitochondrial regions (16S, 12S and control region) performed equally well. The proposed real-time PCR assay can be applied for the accurate confirmation of either lion or tiger DNA that could be used by law enforcement agencies around the world as a tool to monitor illegal trade of tiger bones.