A streamlined, bi-organelle, multiplex PCR approach to species identification:: Application to global conservation and trade monitoring of the great white shark, Carcharodon carcharias

The great white shark, Carcharodon carcharias, is the most widely protected elasmobranch in the world, and is classified as Vulnerable by the IUCN and listed on Appendix III of CITES. Monitoring of trade in white shark products and enforcement of harvest and trade prohibitions is problematic, however, in large part due to difficulties in identifying marketed shark parts (e. g., dried fins, meat and processed carcasses) to species level. To address these conservation and management problems, we have developed a rapid, molecular diagnostic assay based on species-specific PCR primer design for accurate identification of white shark body parts, including dried fins. The assay is novel in several respects: It employs a multiplex PCR assay utilizing both nuclear (ribosomal internal transcribed spacer 2) and mitochondrial (cytochrome b) loci simultaneously to achieve a highly robust measure of diagnostic accuracy; it is very sensitive, detecting the presence of white shark DNA in a mixture of genomic DNAs from up to ten different commercially fished shark species pooled together in a single PCR tube; and it successfully identifies white shark DNA from globally distributed animals. In addition to its utility for white shark trade monitoring and conservation applications, this highly streamlined, bi-organelle, multiplex PCR assay may prove useful as a general model for the design of genetic assays aimed at detecting body parts from other protected and threatened species.