Rapid Detection of Prunus Necrotic Ringspot Virus by Reverse Transcription-cross-priming Amplification Coupled with Nucleic Acid Test Strip Cassette

Sci Rep. 2017 Nov 23;7(1):16175. doi: 10.1038/s41598-017-16536-6.

Abstract

Prunus necrotic ringspot virus (PNRSV) is one of the most devastating viruses to Prunus spp. In this study, we developed a diagnostic system RT-CPA-NATSC, wherein reverse transcription-cross-priming amplification (RT-CPA) is coupled with nucleic acid test strip cassette (NATSC), a vertical flow (VF) visualization, for PNRSV detection. The RT-CPA-NATSC assay targets the encoding gene of the PNRSV coat protein with a limit of detection of 72 copies per reaction and no cross-reaction with the known Prunus pathogenic viruses and viroids, demonstrating high sensitivity and specificity. The reaction is performed on 60 °C and can be completed less than 90 min with the prepared template RNA. Field sample test confirmed the reliability of RT-CPA-NATSC, indicating the potential application of this simple and rapid detection method in routine test of PNRSV.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Cross-Priming / genetics*
  • Cucumis sativus / virology*
  • Ilarvirus / genetics*
  • Ilarvirus / isolation & purification
  • Nucleic Acid Amplification Techniques / methods*
  • Nucleic Acids / genetics
  • Plant Leaves / virology
  • Reverse Transcription / genetics*

Substances

  • Nucleic Acids

Supplementary concepts

  • Prunus necrotic ringspot virus