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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...
Autores principales: | , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Public Library of Science
2014
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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. |
format | Online Article Text |
id | pubmed-4133268 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2014 |
publisher | Public Library of Science |
record_format | MEDLINE/PubMed |
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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