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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Main Authors: Ouso, D.O., Otiende, M.Y., Jeneby, M.M., Oundo, J.W., Bargul, J.L., Miller, S.E., Wambua, Lillian, Villinger, J.
Format: Journal Article
Language:Inglés
Published: Springer 2020
Subjects:
Online Access:https://hdl.handle.net/10568/108124
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author Ouso, D.O.
Otiende, M.Y.
Jeneby, M.M.
Oundo, J.W.
Bargul, J.L.
Miller, S.E.
Wambua, Lillian
Villinger, J.
author_browse Bargul, J.L.
Jeneby, M.M.
Miller, S.E.
Otiende, M.Y.
Oundo, J.W.
Ouso, D.O.
Villinger, J.
Wambua, Lillian
author_facet Ouso, D.O.
Otiende, M.Y.
Jeneby, M.M.
Oundo, J.W.
Bargul, J.L.
Miller, S.E.
Wambua, Lillian
Villinger, J.
author_sort Ouso, D.O.
collection Repository of Agricultural Research Outputs (CGSpace)
description 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.
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spelling CGSpace1081242025-03-11T12:14:31Z Three-gene PCR and high-resolution melting analysis for differentiating vertebrate species mitochondrial DNA for biodiversity research and complementing forensic surveillance Ouso, D.O. Otiende, M.Y. Jeneby, M.M. Oundo, J.W. Bargul, J.L. Miller, S.E. Wambua, Lillian Villinger, J. biodiversity research pcr wildlife forensic sciences 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. 2020-03-16 2020-05-01T12:49:14Z 2020-05-01T12:49:14Z Journal Article https://hdl.handle.net/10568/108124 en Open Access Springer Ouso, D.O., Otiende, M.Y., Jeneby, M.M., Oundo, J.W., Bargul, J.L., Miller, S.E., Wambua, L. and Villinger, J. 2020. Three-gene PCR and high-resolution melting analysis for differentiating vertebrate species mitochondrial DNA for biodiversity research and complementing forensic surveillance. Scientific Reports 10: 4741.
spellingShingle biodiversity
research
pcr
wildlife
forensic sciences
Ouso, D.O.
Otiende, M.Y.
Jeneby, M.M.
Oundo, J.W.
Bargul, J.L.
Miller, S.E.
Wambua, Lillian
Villinger, J.
Three-gene PCR and high-resolution melting analysis for differentiating vertebrate species mitochondrial DNA for biodiversity research and complementing forensic surveillance
title Three-gene PCR and high-resolution melting analysis for differentiating vertebrate species mitochondrial DNA for biodiversity research and complementing forensic surveillance
title_full Three-gene PCR and high-resolution melting analysis for differentiating vertebrate species mitochondrial DNA for biodiversity research and complementing forensic surveillance
title_fullStr Three-gene PCR and high-resolution melting analysis for differentiating vertebrate species mitochondrial DNA for biodiversity research and complementing forensic surveillance
title_full_unstemmed Three-gene PCR and high-resolution melting analysis for differentiating vertebrate species mitochondrial DNA for biodiversity research and complementing forensic surveillance
title_short Three-gene PCR and high-resolution melting analysis for differentiating vertebrate species mitochondrial DNA for biodiversity research and complementing forensic surveillance
title_sort three gene pcr and high resolution melting analysis for differentiating vertebrate species mitochondrial dna for biodiversity research and complementing forensic surveillance
topic biodiversity
research
pcr
wildlife
forensic sciences
url https://hdl.handle.net/10568/108124
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