A streamlined, bi-organelle, multiplex PCR approach to species identification: Application to global conservation and trade monitoring of the great white shark, Carcharodon carcharias |
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Authors: | Demian D. Chapman Debra L. Abercrombie Christophe J. Douady Ellen K. Pikitch Michael J. Stanhopen Mahmood S. Shivji |
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Affiliation: | (1) Guy Harvey Research Institute, Oceanographic Center, Nova Southeastern University, 8000 North Ocean Drive, Dania Beach, FL, 33004, U.S.A;(2) Biology and Biochemistry, The Queen's University, 97 Lisburn Rd., Belfast, N. Ireland, UK;;(3) Present address: Department of Biochemistry and Molecular Biology, Sir Charles Tupper Medical Building, Dalhousie University, Halifax, Nova Scotia, B3H 4H7, Canada;(4) Wildlife Conservation Society, 2300 Southern Boulevard, Bronx, New York, 10460, U.S.A;(5) Biology and Biochemistry, The Queen's, University, 97 Lisburn Rd., Belfast, N. Ireland, UK;;(6) Present address: Evolutionary Bioinformatics, GlaxoSmithKline, 1250 South Collegeville Road, UP1345, Collegeville, PA, 19426-0989, U.S.A |
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Abstract: | The great white shark, Carcharodoncarcharias, is the most widely protectedelasmobranch in the world, and is classified asVulnerable by the IUCN and listed on AppendixIII of CITES. Monitoring of trade in whiteshark products and enforcement of harvest andtrade prohibitions is problematic, however, inlarge part due to difficulties in identifyingmarketed shark parts (e.g., dried fins, meatand processed carcasses) to species level. Toaddress these conservation and managementproblems, we have developed a rapid, moleculardiagnostic assay based on species-specific PCRprimer design for accurate identification ofwhite shark body parts, including dried fins. The assay is novel in several respects: Itemploys a multiplex PCR assay utilizing bothnuclear (ribosomal internal transcribed spacer2) and mitochondrial (cytochrome b) locisimultaneously to achieve a highly robustmeasure of diagnostic accuracy; it is verysensitive, detecting the presence of whiteshark DNA in a mixture of genomic DNAs from upto ten different commercially fished sharkspecies pooled together in a single PCR tube;and it successfully identifies white shark DNAfrom globally distributed animals. Inaddition to its utility for white shark trademonitoring and conservation applications, thishighly streamlined, bi-organelle, multiplex PCRassay may prove useful as a general model forthe design of genetic assays aimed at detectingbody parts from other protected and threatenedspecies. |
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Keywords: | great white sharks multiplex PCR shark conservation species identification |
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