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Comparison of a loop-mediated isothermal amplification for orf virus with quantitative real-time PCR

BACKGROUND: Orf virus (ORFV) causes orf (also known as contagious ecthyma or contagious papular dermatitis), a severe infectious skin disease in goats, sheep and other ruminants. Therefore, a rapid, highly specific and accurate method for the diagnosis of ORFV infections is essential to ensure that...

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Autores principales: Wang, Guangxiang, Shang, Youjun, Wang, Yanhua, Tian, Hong, Liu, Xiangtao
Formato: Online Artículo Texto
Lenguaje:English
Publicado: BioMed Central 2013
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3651318/
https://www.ncbi.nlm.nih.gov/pubmed/23634981
http://dx.doi.org/10.1186/1743-422X-10-138
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author Wang, Guangxiang
Shang, Youjun
Wang, Yanhua
Tian, Hong
Liu, Xiangtao
author_facet Wang, Guangxiang
Shang, Youjun
Wang, Yanhua
Tian, Hong
Liu, Xiangtao
author_sort Wang, Guangxiang
collection PubMed
description BACKGROUND: Orf virus (ORFV) causes orf (also known as contagious ecthyma or contagious papular dermatitis), a severe infectious skin disease in goats, sheep and other ruminants. Therefore, a rapid, highly specific and accurate method for the diagnosis of ORFV infections is essential to ensure that the appropriate treatments are administered and to reduce economic losses. METHODS: A loop-mediated isothermal amplification (LAMP) assay based on the identification of the F1L gene was developed for the specific detection of ORFV infections. The sensitivity and specificity of the LAMP assay were evaluated, and the effectiveness of this method was compared with that of real-time PCR. RESULTS: The sensitivity of this assay was determined to be 10 copies of a standard plasmid. Furthermore, no cross-reactivity was found with either capripox virus or FMDV. The LAMP and real-time PCR assays were both able to detect intracutaneous- and cohabitation-infection samples, with a concordance of 97.83%. LAMP demonstrated a sensitivity of 89.13%. CONCLUSION: The LAMP assay is a highly efficient and practical method for detecting ORFV infection. This LAMP method shows great potential for monitoring the prevalence of orf, and it could prove to be a powerful supplemental tool for current diagnostic methods.
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spelling pubmed-36513182013-05-11 Comparison of a loop-mediated isothermal amplification for orf virus with quantitative real-time PCR Wang, Guangxiang Shang, Youjun Wang, Yanhua Tian, Hong Liu, Xiangtao Virol J Research BACKGROUND: Orf virus (ORFV) causes orf (also known as contagious ecthyma or contagious papular dermatitis), a severe infectious skin disease in goats, sheep and other ruminants. Therefore, a rapid, highly specific and accurate method for the diagnosis of ORFV infections is essential to ensure that the appropriate treatments are administered and to reduce economic losses. METHODS: A loop-mediated isothermal amplification (LAMP) assay based on the identification of the F1L gene was developed for the specific detection of ORFV infections. The sensitivity and specificity of the LAMP assay were evaluated, and the effectiveness of this method was compared with that of real-time PCR. RESULTS: The sensitivity of this assay was determined to be 10 copies of a standard plasmid. Furthermore, no cross-reactivity was found with either capripox virus or FMDV. The LAMP and real-time PCR assays were both able to detect intracutaneous- and cohabitation-infection samples, with a concordance of 97.83%. LAMP demonstrated a sensitivity of 89.13%. CONCLUSION: The LAMP assay is a highly efficient and practical method for detecting ORFV infection. This LAMP method shows great potential for monitoring the prevalence of orf, and it could prove to be a powerful supplemental tool for current diagnostic methods. BioMed Central 2013-05-01 /pmc/articles/PMC3651318/ /pubmed/23634981 http://dx.doi.org/10.1186/1743-422X-10-138 Text en Copyright © 2013 Wang et al.; licensee BioMed Central Ltd. http://creativecommons.org/licenses/by/2.0 This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/2.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
spellingShingle Research
Wang, Guangxiang
Shang, Youjun
Wang, Yanhua
Tian, Hong
Liu, Xiangtao
Comparison of a loop-mediated isothermal amplification for orf virus with quantitative real-time PCR
title Comparison of a loop-mediated isothermal amplification for orf virus with quantitative real-time PCR
title_full Comparison of a loop-mediated isothermal amplification for orf virus with quantitative real-time PCR
title_fullStr Comparison of a loop-mediated isothermal amplification for orf virus with quantitative real-time PCR
title_full_unstemmed Comparison of a loop-mediated isothermal amplification for orf virus with quantitative real-time PCR
title_short Comparison of a loop-mediated isothermal amplification for orf virus with quantitative real-time PCR
title_sort comparison of a loop-mediated isothermal amplification for orf virus with quantitative real-time pcr
topic Research
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3651318/
https://www.ncbi.nlm.nih.gov/pubmed/23634981
http://dx.doi.org/10.1186/1743-422X-10-138
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