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Development of real-time PCR assay for specific detection of cowpox virus

BACKGROUND: The number of recorded human cowpox cases are recently increasing. The symptoms caused by cowpox virus (CPXV) in a number of human cases are close to the symptoms characteristic of the orthopoxviral human infections caused by monkeypox or smallpox (variola) viruses. Any rapid and reliabl...

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Autores principales: Gavrilova, Elena V., Shcherbakov, Dmitrii N., Maksyutov, Rinat A., Shchelkunov, Sergei N.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Elsevier B.V. 2010
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9628739/
https://www.ncbi.nlm.nih.gov/pubmed/20594906
http://dx.doi.org/10.1016/j.jcv.2010.06.003
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author Gavrilova, Elena V.
Shcherbakov, Dmitrii N.
Maksyutov, Rinat A.
Shchelkunov, Sergei N.
author_facet Gavrilova, Elena V.
Shcherbakov, Dmitrii N.
Maksyutov, Rinat A.
Shchelkunov, Sergei N.
author_sort Gavrilova, Elena V.
collection PubMed
description BACKGROUND: The number of recorded human cowpox cases are recently increasing. The symptoms caused by cowpox virus (CPXV) in a number of human cases are close to the symptoms characteristic of the orthopoxviral human infections caused by monkeypox or smallpox (variola) viruses. Any rapid and reliable real-time PCR method for distinguishing cowpox from smallpox and monkeypox is yet absent. OBJECTIVES: The aim of this study was to develop a quick and reliable real-time TaqMan PCR assay for specific detection of cowpox virus and to determine the sensitivity and specificity of this method. STUDY DESIGN: Based on aligned nucleotide sequences of orthopoxviruses, we found a virus-specific region in the CPXV genome and selected the oligonucleotide primers and hybridization probe within this region. The specificity of the developed method was tested using a panel of various orthopoxvirus (OPV) DNAs. The sensitivity was determined using the recombinant plasmid carrying a fragment of CPXV DNA and genomic DNA of the CPXV strain GRI-90. RESULTS: The analytical specificity of this method was determined using DNAs of 17 strains of four OPV species pathogenic for humans and amounted to 100%. The method allows 6 copies of plasmid DNA and 20 copies of CPXV DNA in the reaction mixture to be detected. CONCLUSION: A quick and reliable TaqMan PCR assay providing for a highly sensitive and specific detection of CPXV DNA was developed.
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spelling pubmed-96287392022-11-03 Development of real-time PCR assay for specific detection of cowpox virus Gavrilova, Elena V. Shcherbakov, Dmitrii N. Maksyutov, Rinat A. Shchelkunov, Sergei N. J Clin Virol Article BACKGROUND: The number of recorded human cowpox cases are recently increasing. The symptoms caused by cowpox virus (CPXV) in a number of human cases are close to the symptoms characteristic of the orthopoxviral human infections caused by monkeypox or smallpox (variola) viruses. Any rapid and reliable real-time PCR method for distinguishing cowpox from smallpox and monkeypox is yet absent. OBJECTIVES: The aim of this study was to develop a quick and reliable real-time TaqMan PCR assay for specific detection of cowpox virus and to determine the sensitivity and specificity of this method. STUDY DESIGN: Based on aligned nucleotide sequences of orthopoxviruses, we found a virus-specific region in the CPXV genome and selected the oligonucleotide primers and hybridization probe within this region. The specificity of the developed method was tested using a panel of various orthopoxvirus (OPV) DNAs. The sensitivity was determined using the recombinant plasmid carrying a fragment of CPXV DNA and genomic DNA of the CPXV strain GRI-90. RESULTS: The analytical specificity of this method was determined using DNAs of 17 strains of four OPV species pathogenic for humans and amounted to 100%. The method allows 6 copies of plasmid DNA and 20 copies of CPXV DNA in the reaction mixture to be detected. CONCLUSION: A quick and reliable TaqMan PCR assay providing for a highly sensitive and specific detection of CPXV DNA was developed. Elsevier B.V. 2010-09 2010-07-01 /pmc/articles/PMC9628739/ /pubmed/20594906 http://dx.doi.org/10.1016/j.jcv.2010.06.003 Text en Copyright © 2010 Elsevier B.V. All rights reserved. Elsevier has created a Monkeypox Information Center (https://www.elsevier.com/connect/monkeypox-information-center) in response to the declared public health emergency of international concern, with free information in English on the monkeypox virus. The Monkeypox Information Center is hosted on Elsevier Connect, the company's public news and information website. Elsevier hereby grants permission to make all its monkeypox related research that is available on the Monkeypox Information Center - including this research content - immediately available in publicly funded repositories, with rights for unrestricted research re-use and analyses in any form or by any means with acknowledgement of the original source. These permissions are granted for free by Elsevier for as long as the Monkeypox Information Center remains active.
spellingShingle Article
Gavrilova, Elena V.
Shcherbakov, Dmitrii N.
Maksyutov, Rinat A.
Shchelkunov, Sergei N.
Development of real-time PCR assay for specific detection of cowpox virus
title Development of real-time PCR assay for specific detection of cowpox virus
title_full Development of real-time PCR assay for specific detection of cowpox virus
title_fullStr Development of real-time PCR assay for specific detection of cowpox virus
title_full_unstemmed Development of real-time PCR assay for specific detection of cowpox virus
title_short Development of real-time PCR assay for specific detection of cowpox virus
title_sort development of real-time pcr assay for specific detection of cowpox virus
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9628739/
https://www.ncbi.nlm.nih.gov/pubmed/20594906
http://dx.doi.org/10.1016/j.jcv.2010.06.003
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