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Microarray for Identification of the Chiropteran Host Species of Rabies Virus in Canada

Species identification through genetic barcoding can augment traditional taxonomic methods, which rely on morphological features of the specimen. Such approaches are especially valuable when specimens are in poor condition or comprise very limited material, a situation that often applies to chiropte...

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Autores principales: Lung, Oliver, Nadin-Davis, Susan, Fisher, Mathew, Erickson, Anthony, Knowles, M. Kimberly, Furukawa-Stoffer, Tara, Ambagala, Aruna
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2013
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5003475/
https://www.ncbi.nlm.nih.gov/pubmed/27605186
http://dx.doi.org/10.3390/microarrays2020153
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author Lung, Oliver
Nadin-Davis, Susan
Fisher, Mathew
Erickson, Anthony
Knowles, M. Kimberly
Furukawa-Stoffer, Tara
Ambagala, Aruna
author_facet Lung, Oliver
Nadin-Davis, Susan
Fisher, Mathew
Erickson, Anthony
Knowles, M. Kimberly
Furukawa-Stoffer, Tara
Ambagala, Aruna
author_sort Lung, Oliver
collection PubMed
description Species identification through genetic barcoding can augment traditional taxonomic methods, which rely on morphological features of the specimen. Such approaches are especially valuable when specimens are in poor condition or comprise very limited material, a situation that often applies to chiropteran (bat) specimens submitted to the Canadian Food Inspection Agency for rabies diagnosis. Coupled with phenotypic plasticity of many species and inconclusive taxonomic keys, species identification using only morphological traits can be challenging. In this study, a microarray assay with associated PCR of the mitochondrial cytochrome c oxidase subunit I (COI) gene was developed for differentiation of 14 bat species submitted to the Canadian Food Inspection Agency from 1985–2012 for rabies diagnosis. The assay was validated with a reference collection of DNA from 153 field samples, all of which had been barcoded previously. The COI gene from 152 samples which included multiple specimens of each target species were successfully amplified by PCR and accurately identified by the microarray. One sample that was severely decomposed failed to amplify with PCR primers developed in this study, but amplified weakly after switching to alternate primers and was accurately typed by the microarray. Thus, the chiropteran microarray was able to accurately differentiate between the 14 species of Canadian bats targeted. This PCR and microarray assay would allow unequivocal identification to species of most, if not all, bat specimens submitted for rabies diagnosis in Canada.
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spelling pubmed-50034752016-09-06 Microarray for Identification of the Chiropteran Host Species of Rabies Virus in Canada Lung, Oliver Nadin-Davis, Susan Fisher, Mathew Erickson, Anthony Knowles, M. Kimberly Furukawa-Stoffer, Tara Ambagala, Aruna Microarrays (Basel) Article Species identification through genetic barcoding can augment traditional taxonomic methods, which rely on morphological features of the specimen. Such approaches are especially valuable when specimens are in poor condition or comprise very limited material, a situation that often applies to chiropteran (bat) specimens submitted to the Canadian Food Inspection Agency for rabies diagnosis. Coupled with phenotypic plasticity of many species and inconclusive taxonomic keys, species identification using only morphological traits can be challenging. In this study, a microarray assay with associated PCR of the mitochondrial cytochrome c oxidase subunit I (COI) gene was developed for differentiation of 14 bat species submitted to the Canadian Food Inspection Agency from 1985–2012 for rabies diagnosis. The assay was validated with a reference collection of DNA from 153 field samples, all of which had been barcoded previously. The COI gene from 152 samples which included multiple specimens of each target species were successfully amplified by PCR and accurately identified by the microarray. One sample that was severely decomposed failed to amplify with PCR primers developed in this study, but amplified weakly after switching to alternate primers and was accurately typed by the microarray. Thus, the chiropteran microarray was able to accurately differentiate between the 14 species of Canadian bats targeted. This PCR and microarray assay would allow unequivocal identification to species of most, if not all, bat specimens submitted for rabies diagnosis in Canada. MDPI 2013-05-31 /pmc/articles/PMC5003475/ /pubmed/27605186 http://dx.doi.org/10.3390/microarrays2020153 Text en © 2013 by the authors; licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution license (http://creativecommons.org/licenses/by/3.0/).
spellingShingle Article
Lung, Oliver
Nadin-Davis, Susan
Fisher, Mathew
Erickson, Anthony
Knowles, M. Kimberly
Furukawa-Stoffer, Tara
Ambagala, Aruna
Microarray for Identification of the Chiropteran Host Species of Rabies Virus in Canada
title Microarray for Identification of the Chiropteran Host Species of Rabies Virus in Canada
title_full Microarray for Identification of the Chiropteran Host Species of Rabies Virus in Canada
title_fullStr Microarray for Identification of the Chiropteran Host Species of Rabies Virus in Canada
title_full_unstemmed Microarray for Identification of the Chiropteran Host Species of Rabies Virus in Canada
title_short Microarray for Identification of the Chiropteran Host Species of Rabies Virus in Canada
title_sort microarray for identification of the chiropteran host species of rabies virus in canada
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5003475/
https://www.ncbi.nlm.nih.gov/pubmed/27605186
http://dx.doi.org/10.3390/microarrays2020153
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