Multiplex real-time PCR assay to detect illegal trade of CITES-listed shark species

The Convention on International Trade in Endangered Species of Wild Fauna and Flora (CITES) is a multilateral environmental agreement to ensure that the international trade of threatened species is either prohibited (Appendix I listed species) or being conducted legally, sustainably, and transparently (Appendix II listed species). Twelve threatened shark species exploited for their fins, meat, and other products have been listed under CITES Appendix II. Sharks are often traded in high volumes, some of their products are visually indistinguishable, and most importing/exporting nations have limited capacity to detect illicit trade and enforce the regulations. High volume shipments often must be screened after only a short period of detainment (e.g., a maximum of 24 hours), which together with costs and capacity issues have limited the use of DNA approaches to identify illicit trade. Here, we present a reliable, field-based, fast (<4 hours), and cost effective ($0.94 USD per sample) multiplex realtime PCR protocol capable of detecting nine of the twelve sharks listed under CITES in a single reaction. This approach facilitates detection of illicit trade, with positive results providing probable cause to detain shipments for more robust forensic analysis. We also provide evidence of its application in real law enforcement scenarios in Hong Kong. Adoption of this approach can help parties meet their CITES requirements, avoiding potential international trade sanctions in the future.