Distinguishing Anuran species by high‐resolution melting analysis of the COI barcode (COI‐HRM)

Everman, Steven; Wang, Shiao Y.

doi:10.1002/ece3.5808

Cited by 7 publications

(8 citation statements)

References 21 publications

(27 reference statements)

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“…The distinction between pairs or groups of sometimes strikingly similar species is often a key element when tackling biological, ecological or conservational research questions [ 24 ]. In the recent past, molecular biological methods have increasingly been used to aid in species assignment, albeit often requiring a substantial amount of infrastructure.…”

Section: Resultsmentioning

confidence: 99%

“…2 c), thus allowing for the assignment of the 13 larvae to either of the two species. The significant outcome of these tests indicates that shape, amplitude and melting peak do not just vary by chance [ 24 ]. The sensitivity of this method is known to account for single nucleotide differences [ 8 ].…”

Section: Resultsmentioning

confidence: 99%

“…The distinction between pairs or groups of sometimes strikingly similar species is often a key element when tackling biological, ecological or conservational research questions [24].…”

Section: Resultsmentioning

confidence: 99%

“…The T m from all samples grouped by taxa were compared with a two-sided Welch t-test using a 95% confidence interval and 10,000 bootstrap replicates. Euclidean distance comparison of melt curves followed [ 24 ]. p-values below 0.05 were considered significant.…”

Section: Methodsmentioning

confidence: 99%

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Discriminating larvae of two syntopic Cychramus species (Coleoptera, Nitidulidae) by means of bar-HRM analysis

Zangl

Oberreiter

Huss³

et al. 2020

Mol Biol Rep

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Molecular genetic methods are increasingly used to supplement or substitute classical morphology-based species identification. Here, we employ a COI mini-barcode coupled high-resolution melting analysis to quickly, cost-efficiently and reliably determine larvae of two closely related Cychramus (Coleoptera, Nitidulidae) species. Euclidean distance comparison (p < 0.01) and a Welch t-test of the melting point temperatures (p < 0.01) provide highly significant statistical evidence for species specific differences in melting and fluorescence curves, thus allowing the assignment of larvae to either of the two species. This protocol serves as a fast, low-cost and low-tech method to discriminate between pairs or groups of closely related species and can be adapted and applied to various ecological research questions.

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Section: Resultsmentioning

confidence: 99%

Section: Resultsmentioning

confidence: 99%

“…The distinction between pairs or groups of sometimes strikingly similar species is often a key element when tackling biological, ecological or conservational research questions [24].…”

Section: Resultsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

See 2 more Smart Citations

Discriminating larvae of two syntopic Cychramus species (Coleoptera, Nitidulidae) by means of bar-HRM analysis

Zangl

Oberreiter

Huss³

et al. 2020

Mol Biol Rep

View full text Add to dashboard Cite

show abstract

“…The amplicon melting temperatures (T m ) and specific melt curve shapes are dependent on DNA complementarity, the order of DNA bases, G-C content, and amplicon length. PCR-HRM has been used with a number of genes to identify species among diverse viruses 26 , bacteria 27 , malaria Plasmodium 28 , mosquitoes 29 and their bloodmeals 30 , plant products 31 , and animals within discrete families 32,33 , as well as human individuals 34 . It is quick (occurs within a real-time PCR machine immediately after PCR) and does not require downstream analyses such as gel electrophoresis to determine amplification success or PCR-product sequencing to differentiate sequence variants.…”

mentioning

confidence: 99%

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

Ouso

Otiende

Jeneby

et al. 2020

Sci Rep

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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. Unsustainable hunting, consumption, and sale of bushmeat in Africa contribute immensely to the decline of threatened wild animal species. The global bushmeat trade is valued at several billion US dollars. Up to 270 tons of bushmeat were flown into Europe through a single airport in 2010 from Africa 1. While this is a major crisis for wildlife in central and western Africa, it is a growing concern in eastern and southern Africa 2. Efforts to regulate or prevent illegal wildlife trade depends on accurate, efficient, and sustainable tools for species identification of confiscated and surveillance samples. Illegal wildlife trade is mainly fuelled by the need for diet and income supplementation 3. The consequences of direct human contact with bushmeat have been severe. A classic example of disease originating from or harboured by bushmeat is Ebola virus disease, which has infected humans upon contact with infected wild animals such as fruit bats, nonhuman primates, and forest antelopes 4,5. The impact of bushmeat hunting on animal populations can also be severe 6. Many favoured wild animal species for bushmeat are already endangered, some close to extinction 7. There are also flow-on effects to the ecosystems 8 and tourism.

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Genotypic analysis of SLC4A1 A858D mutation in Indian population associated with distal renal tubular Acidosis (dRTA) coupled with hemolytic anemia

Kedar

2021

Gene

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Distinguishing Anuran species by high‐resolution melting analysis of the COI barcode (COI‐HRM)

Cited by 7 publications

References 21 publications

Discriminating larvae of two syntopic Cychramus species (Coleoptera, Nitidulidae) by means of bar-HRM analysis

Discriminating larvae of two syntopic Cychramus species (Coleoptera, Nitidulidae) by means of bar-HRM analysis

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

Genotypic analysis of SLC4A1 A858D mutation in Indian population associated with distal renal tubular Acidosis (dRTA) coupled with hemolytic anemia

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