A new assay for identifying endangered species in Traditional East Asian Medicine

Despite renewed public interest, education, research and legislation, Traditional East Asian Medication (TEAM) continues to incorporate animal parts from endangered species and can be obtained openly in many countries. Products sold as having medicinal properties include plasters, pills, ointments, tonic wine and others. Due to the low population levels of some endangered species other more common species may be substituted or very small amounts may be used. This results in low levels of DNA which can be difficult to identify. Most species identification tests rely on sequence comparison with a known reference sample. Most TEAM contains mixtures of several different species making this technique unsuitable. We describe a novel single step assay to rapidly and simultaneously identify rhino, tiger, bear, leopard, pangolin, musk deer and several non-endangered mammals often substituted in TEAM. The test targets the mitochondrial genome to amplify species-specific fragments that can be separated easily using a genetic analyzer. Each fragment is of a different size so that none can be confused. The specificity of each primer pair allows for species identification to be made even if a mixture of several species is present. Further, if more than one of the target species is present then all the species will be amplified and identified simultaneously. The test is sensitive to very low levels of DNA equating to several hundred mitochondrial copies (a fraction of single cell).