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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Published in:PloS one 2014-08, Vol.9 (8), p.e104667-e104667
Main Authors: Xia, Xiaoming, Yu, Yongxin, Weidmann, Manfred, Pan, Yingjie, Yan, Shuling, Wang, Yongjie
Format: Article
Language:English
Subjects:
Amplification
Animals
Aquaculture
Aquaculture industry
Assaying
Base Sequence
Biochemistry
Biology and Life Sciences
Comparative analysis
Crustacea
Crustacea - virology
Crustaceans
Deoxyribonucleic acid
DNA
DNA Primers
DNA, Viral - analysis
Economic impact
Engineering research
Farms
Food science
Genetic testing
Health aspects
Laboratories
Limit of Detection
Plasmids
Polymerase chain reaction
Quality
Real time
Real-Time Polymerase Chain Reaction - methods
Recombinase
Reproducibility of Results
Risk assessment
Seafood industry
Sensitivity analysis
Shellfish
Virology
Viruses
White spot syndrome
White spot syndrome virus 1 - genetics
White spot syndrome virus 1 - isolation & purification
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Weidmann, Manfred
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Yan, Shuling
Wang, Yongjie
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.
doi_str_mv 10.1371/journal.pone.0104667
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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. 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subjects Amplification
Animals
Aquaculture
Aquaculture industry
Assaying
Base Sequence
Biochemistry
Biology and Life Sciences
Comparative analysis
Crustacea
Crustacea - virology
Crustaceans
Deoxyribonucleic acid
DNA
DNA Primers
DNA, Viral - analysis
Economic impact
Engineering research
Farms
Food science
Genetic testing
Health aspects
Laboratories
Limit of Detection
Plasmids
Polymerase chain reaction
Quality
Real time
Real-Time Polymerase Chain Reaction - methods
Recombinase
Reproducibility of Results
Risk assessment
Seafood industry
Sensitivity analysis
Shellfish
Virology
Viruses
White spot syndrome
White spot syndrome virus 1 - genetics
White spot syndrome virus 1 - isolation & purification
title Rapid detection of shrimp white spot syndrome virus by real time, isothermal recombinase polymerase amplification assay
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