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Rapid and High-Throughput pan-Orthopoxvirus Detection and Identification using PCR and Mass Spectrometry

The genus Orthopoxvirus contains several species of related viruses, including the causative agent of smallpox (Variola virus). In addition to smallpox, several other members of the genus are capable of causing human infection, including monkeypox, cowpox, and other zoonotic rodent-borne poxviruses....

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Autores principales: Eshoo, Mark W., Whitehouse, Chris A., Nalca, Aysegul, Zoll, Scott, Ecker, Joseph A., Hall, Thomas A., Pennella, Thuy-Trang D., Duncan, David D., Desai, Anjali, Moradi, Emily K., Rudnick, Karl, Libby, Brian, Ranken, Raymond, Sampath, Rangarajan, Hofstadler, Steven A., Ecker, David J., Blyn, Lawrence B.
Formato: Texto
Lenguaje:English
Publicado: Public Library of Science 2009
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2710004/
https://www.ncbi.nlm.nih.gov/pubmed/19623263
http://dx.doi.org/10.1371/journal.pone.0006342
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author Eshoo, Mark W.
Whitehouse, Chris A.
Nalca, Aysegul
Zoll, Scott
Ecker, Joseph A.
Hall, Thomas A.
Pennella, Thuy-Trang D.
Duncan, David D.
Desai, Anjali
Moradi, Emily K.
Rudnick, Karl
Libby, Brian
Ranken, Raymond
Sampath, Rangarajan
Hofstadler, Steven A.
Ecker, David J.
Blyn, Lawrence B.
author_facet Eshoo, Mark W.
Whitehouse, Chris A.
Nalca, Aysegul
Zoll, Scott
Ecker, Joseph A.
Hall, Thomas A.
Pennella, Thuy-Trang D.
Duncan, David D.
Desai, Anjali
Moradi, Emily K.
Rudnick, Karl
Libby, Brian
Ranken, Raymond
Sampath, Rangarajan
Hofstadler, Steven A.
Ecker, David J.
Blyn, Lawrence B.
author_sort Eshoo, Mark W.
collection PubMed
description The genus Orthopoxvirus contains several species of related viruses, including the causative agent of smallpox (Variola virus). In addition to smallpox, several other members of the genus are capable of causing human infection, including monkeypox, cowpox, and other zoonotic rodent-borne poxviruses. Therefore, a single assay that can accurately identify all orthopoxviruses could provide a valuable tool for rapid broad orthopovirus identification. We have developed a pan-Orthopoxvirus assay for identification of all members of the genus based on four PCR reactions targeting Orthopoxvirus DNA and RNA helicase and polymerase genes. The amplicons are detected using electrospray ionization-mass spectrometry (PCR/ESI-MS) on the Ibis T5000 system. We demonstrate that the assay can detect and identify a diverse collection of orthopoxviruses, provide sub-species information and characterize viruses from the blood of rabbitpox infected rabbits. The assay is sensitive at the stochastic limit of PCR and detected virus in blood containing approximately six plaque-forming units per milliliter from a rabbitpox virus-infected rabbit.
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spelling pubmed-27100042009-07-22 Rapid and High-Throughput pan-Orthopoxvirus Detection and Identification using PCR and Mass Spectrometry Eshoo, Mark W. Whitehouse, Chris A. Nalca, Aysegul Zoll, Scott Ecker, Joseph A. Hall, Thomas A. Pennella, Thuy-Trang D. Duncan, David D. Desai, Anjali Moradi, Emily K. Rudnick, Karl Libby, Brian Ranken, Raymond Sampath, Rangarajan Hofstadler, Steven A. Ecker, David J. Blyn, Lawrence B. PLoS One Research Article The genus Orthopoxvirus contains several species of related viruses, including the causative agent of smallpox (Variola virus). In addition to smallpox, several other members of the genus are capable of causing human infection, including monkeypox, cowpox, and other zoonotic rodent-borne poxviruses. Therefore, a single assay that can accurately identify all orthopoxviruses could provide a valuable tool for rapid broad orthopovirus identification. We have developed a pan-Orthopoxvirus assay for identification of all members of the genus based on four PCR reactions targeting Orthopoxvirus DNA and RNA helicase and polymerase genes. The amplicons are detected using electrospray ionization-mass spectrometry (PCR/ESI-MS) on the Ibis T5000 system. We demonstrate that the assay can detect and identify a diverse collection of orthopoxviruses, provide sub-species information and characterize viruses from the blood of rabbitpox infected rabbits. The assay is sensitive at the stochastic limit of PCR and detected virus in blood containing approximately six plaque-forming units per milliliter from a rabbitpox virus-infected rabbit. Public Library of Science 2009-07-22 /pmc/articles/PMC2710004/ /pubmed/19623263 http://dx.doi.org/10.1371/journal.pone.0006342 Text en Eshoo et al. http://creativecommons.org/licenses/by/4.0/ This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are properly credited.
spellingShingle Research Article
Eshoo, Mark W.
Whitehouse, Chris A.
Nalca, Aysegul
Zoll, Scott
Ecker, Joseph A.
Hall, Thomas A.
Pennella, Thuy-Trang D.
Duncan, David D.
Desai, Anjali
Moradi, Emily K.
Rudnick, Karl
Libby, Brian
Ranken, Raymond
Sampath, Rangarajan
Hofstadler, Steven A.
Ecker, David J.
Blyn, Lawrence B.
Rapid and High-Throughput pan-Orthopoxvirus Detection and Identification using PCR and Mass Spectrometry
title Rapid and High-Throughput pan-Orthopoxvirus Detection and Identification using PCR and Mass Spectrometry
title_full Rapid and High-Throughput pan-Orthopoxvirus Detection and Identification using PCR and Mass Spectrometry
title_fullStr Rapid and High-Throughput pan-Orthopoxvirus Detection and Identification using PCR and Mass Spectrometry
title_full_unstemmed Rapid and High-Throughput pan-Orthopoxvirus Detection and Identification using PCR and Mass Spectrometry
title_short Rapid and High-Throughput pan-Orthopoxvirus Detection and Identification using PCR and Mass Spectrometry
title_sort rapid and high-throughput pan-orthopoxvirus detection and identification using pcr and mass spectrometry
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2710004/
https://www.ncbi.nlm.nih.gov/pubmed/19623263
http://dx.doi.org/10.1371/journal.pone.0006342
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