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Rapid Detection of Shrimp White Spot Syndrome Virus by Real Time, Isothermal Recombinase Polymerase Amplification Assay

White spot syndrome virus (WSSV) causes large economic losses to the shrimp aquaculture industry, and thus far there are no efficient therapeutic treatments available against this lethal virus. In this study, we present the development of a novel real time isothermal recombinase polymerase amplifica...

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Detalles Bibliográficos
Autores principales: Xia, Xiaoming, Yu, Yongxin, Weidmann, Manfred, Pan, Yingjie, Yan, Shuling, Wang, Yongjie
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Public Library of Science 2014
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4133268/
https://www.ncbi.nlm.nih.gov/pubmed/25121957
http://dx.doi.org/10.1371/journal.pone.0104667
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author Xia, Xiaoming
Yu, Yongxin
Weidmann, Manfred
Pan, Yingjie
Yan, Shuling
Wang, Yongjie
author_facet Xia, Xiaoming
Yu, Yongxin
Weidmann, Manfred
Pan, Yingjie
Yan, Shuling
Wang, Yongjie
author_sort Xia, Xiaoming
collection PubMed
description White spot syndrome virus (WSSV) causes large economic losses to the shrimp aquaculture industry, and thus far there are no efficient therapeutic treatments available against this lethal virus. In this study, we present the development of a novel real time isothermal recombinase polymerase amplification (RPA) assay for WSSV detection on a small ESEQuant Tube Scanner device. The RPA sensitivity, specificity and rapidity were evaluated by using a plasmid standard as well as viral and shrimp genomic DNAs. Compared with qPCR, the RPA assay revealed more satisfactory performance. It reached a detection limit up to 10 molecules in 95% of cases as determined by probit analysis of 8 independent experiments within 6.41±0.17 min at 39°C. Consequently, this rapid RPA method has great application potential for field use or point of care diagnostics.
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spelling pubmed-41332682014-08-19 Rapid Detection of Shrimp White Spot Syndrome Virus by Real Time, Isothermal Recombinase Polymerase Amplification Assay Xia, Xiaoming Yu, Yongxin Weidmann, Manfred Pan, Yingjie Yan, Shuling Wang, Yongjie PLoS One Research Article White spot syndrome virus (WSSV) causes large economic losses to the shrimp aquaculture industry, and thus far there are no efficient therapeutic treatments available against this lethal virus. In this study, we present the development of a novel real time isothermal recombinase polymerase amplification (RPA) assay for WSSV detection on a small ESEQuant Tube Scanner device. The RPA sensitivity, specificity and rapidity were evaluated by using a plasmid standard as well as viral and shrimp genomic DNAs. Compared with qPCR, the RPA assay revealed more satisfactory performance. It reached a detection limit up to 10 molecules in 95% of cases as determined by probit analysis of 8 independent experiments within 6.41±0.17 min at 39°C. Consequently, this rapid RPA method has great application potential for field use or point of care diagnostics. Public Library of Science 2014-08-14 /pmc/articles/PMC4133268/ /pubmed/25121957 http://dx.doi.org/10.1371/journal.pone.0104667 Text en © 2014 Xia 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
Xia, Xiaoming
Yu, Yongxin
Weidmann, Manfred
Pan, Yingjie
Yan, Shuling
Wang, Yongjie
Rapid Detection of Shrimp White Spot Syndrome Virus by Real Time, Isothermal Recombinase Polymerase Amplification Assay
title Rapid Detection of Shrimp White Spot Syndrome Virus by Real Time, Isothermal Recombinase Polymerase Amplification Assay
title_full Rapid Detection of Shrimp White Spot Syndrome Virus by Real Time, Isothermal Recombinase Polymerase Amplification Assay
title_fullStr Rapid Detection of Shrimp White Spot Syndrome Virus by Real Time, Isothermal Recombinase Polymerase Amplification Assay
title_full_unstemmed Rapid Detection of Shrimp White Spot Syndrome Virus by Real Time, Isothermal Recombinase Polymerase Amplification Assay
title_short Rapid Detection of Shrimp White Spot Syndrome Virus by Real Time, Isothermal Recombinase Polymerase Amplification Assay
title_sort rapid detection of shrimp white spot syndrome virus by real time, isothermal recombinase polymerase amplification assay
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4133268/
https://www.ncbi.nlm.nih.gov/pubmed/25121957
http://dx.doi.org/10.1371/journal.pone.0104667
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