Rapid detection of shrimp white spot syndrome virus by real time, isothermal recombinase polymerase amplification assay

PLoS One. 2014 Aug 14;9(8):e104667. doi: 10.1371/journal.pone.0104667. eCollection 2014.

Abstract

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.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Base Sequence
  • Crustacea / virology
  • DNA Primers
  • DNA, Viral / analysis
  • Limit of Detection
  • Plasmids
  • Real-Time Polymerase Chain Reaction / methods*
  • Reproducibility of Results
  • White spot syndrome virus 1 / genetics
  • White spot syndrome virus 1 / isolation & purification*

Substances

  • DNA Primers
  • DNA, Viral

Grants and funding

This work was supported by the National Natural Science Foundation of China (No. 41376135), Doctoral Fund of Ministry of Education of China (20133104110006), Innovation Program of Shanghai Municipal Education Commission (14ZZ144), China, and Construction Program of Shanghai Committee of Science and Technology (11DZ2280300), China. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.