A mitochondrial species identification assay for Australian blacktip sharks (Carcharhinus tilstoni, C. limbatus and C. amblyrhynchoides) using real-time PCR and high-resolution melt analysis

Morgan, Jess A. T., Welch, David J., Harry, Alistair V., Street, Raewyn, Broderick, Damien and Ovenden, Jennifer R. (2011) A mitochondrial species identification assay for Australian blacktip sharks (Carcharhinus tilstoni, C. limbatus and C. amblyrhynchoides) using real-time PCR and high-resolution melt analysis. Molecular Ecology Resources, 11 5: 813-819.

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Author Morgan, Jess A. T.
Welch, David J.
Harry, Alistair V.
Street, Raewyn
Broderick, Damien
Ovenden, Jennifer R.
Title A mitochondrial species identification assay for Australian blacktip sharks (Carcharhinus tilstoni, C. limbatus and C. amblyrhynchoides) using real-time PCR and high-resolution melt analysis
Formatted title A mitochondrial species identification assay for Australian blacktip sharks (Carcharhinus tilstoni, C. limbatus and C. amblyrhynchoides) using real-time PCR and high-resolution melt analysis
Journal name Molecular Ecology Resources   Check publisher's open access policy
ISSN 1755-098X
1755-0998
Publication date 2011-09
Sub-type Article (original research)
DOI 10.1111/j.1755-0998.2011.03023.x
Volume 11
Issue 5
Start page 813
End page 819
Total pages 7
Place of publication Oxford, U.K.
Publisher Wiley-Blackwell Publishing
Collection year 2012
Language eng
Formatted abstract Tropical Australian shark fisheries target two morphologically indistinguishable blacktip sharks, the Australian blacktip (Carcharhinus tilstoni) and the common blacktip (C. limbatus). Their relative contributions to northern and eastern Australian coastal fisheries are unclear because of species identification difficulties. The two species differ in their number of precaudal vertebrae, which is difficult and time consuming to obtain in the field. But, the two species can be distinguished genetically with diagnostic mutations in their mitochondrial DNA ND4 gene. A third closely related sister species, the graceful shark C. amblyrhynchoides, can also be distinguished by species-specific mutations in this gene. DNA sequencing is an effective diagnostic tool, but is relatively expensive and time consuming. In contrast, real-time high-resolution melt (HRM) PCR assays are rapid and relatively inexpensive. These assays amplify regions of DNA with species-specific genetic mutations that result in PCR products with unique melt profiles. A real-time HRM PCR species-diagnostic assay (RT-HRM-PCR) has been developed based on the mtDNA ND4 gene for rapid typing of C. tilstoni, C. limbatus and C. amblyrhynchoides. The assay was developed using ND4 sequences from 66 C. tilstoni, 33. C. limbatus and five C. amblyrhynchoides collected from Indonesia and Australian states and territories; Western Australia, the Northern Territory, Queensland and New South Wales. The assay was shown to be 100% accurate on 160 unknown blacktip shark tissue samples by full mtDNA ND4 sequencing.
Keyword Australian blacktip shark
Common blacktip shark
Graceful shark
Mitochondrial DNA ND4 region
Phylogenetic-relationships
Multiplex PCR
DNA
Chondrichthyans
Conservation
Q-Index Code C1
Q-Index Status Confirmed Code
Institutional Status UQ

Document type: Journal Article
Sub-type: Article (original research)
Collections: Official 2012 Collection
Queensland Alliance for Agriculture and Food Innovation
 
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