Three-gene PCR and high-resolution melting analysis for differentiating vertebrate species mitochondrial DNA for biodiversity research and complementing forensic surveillance

Reliable molecular identification of vertebrate species from morphologically unidentifiable tissue is critical for the prosecution of illegally-traded wildlife products, conservation-based biodiversity research, and identification of blood-meal hosts of hematophagous invertebrates. However, forensic...

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Bibliographic Details
Published in:Scientific reports 2020-03, Vol.10 (1), p.4741-4741, Article 4741
Main Authors: Ouso, Daniel O, Otiende, Moses Y, Jeneby, Maamun M, Oundo, Joseph W, Bargul, Joel L, Miller, Scott E, Wambua, Lillian, Villinger, Jandouwe
Format: Article
Language:English
Subjects:
Animals
Animals, Wild - genetics
Biodiversity
Conservation of Natural Resources
Cytochrome
Cytochrome b
Cytochrome oxidase I
Cytochromes b - genetics
DNA barcoding
DNA, Mitochondrial - genetics
Electron Transport Complex IV - genetics
Elephants - genetics
Forensic Genetics - methods
Forensic science
Genetic analysis
Giraffes - genetics
High-Throughput Screening Assays - methods
Invertebrates
Kenya
Loxodonta africana
Loxodonta cyclotis
Meat
Mitochondria - enzymology
Mitochondrial DNA
Polymerase chain reaction
Polymerase Chain Reaction - methods
RNA, Ribosomal, 16S - genetics
rRNA 16S
Savannahs
Species
Species Specificity
Surveillance
Trade
Vertebrates - genetics
Wildlife conservation
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Summary:Reliable molecular identification of vertebrate species from morphologically unidentifiable tissue is critical for the prosecution of illegally-traded wildlife products, conservation-based biodiversity research, and identification of blood-meal hosts of hematophagous invertebrates. However, forensic identification of vertebrate tissue relies on sequencing of the mitochondrial cytochrome oxidase I (COI) 'barcode' gene, which remains costly for purposes of screening large numbers of unknown samples during routine surveillance. Here, we adapted a rapid, low-cost approach to differentiate 10 domestic and 24 wildlife species that are common in the East African illegal wildlife products trade based on their unique high-resolution melting profiles from COI, cytochrome b, and 16S ribosomal RNA gene PCR products. Using the approach, we identified (i) giraffe among covertly sampled meat from Kenyan butcheries, and (ii) forest elephant mitochondrial sequences among savannah elephant reference samples. This approach is being adopted for high-throughput pre-screening of potential bushmeat samples in East African forensic science pipelines.
ISSN:2045-2322
2045-2322
DOI:10.1038/s41598-020-61600-3