Reverse transcription-recombinase polymerase amplification combined with lateral flow strip for detection of rice black-streaked dwarf virus in plants

J Virol Methods. 2019 Jan:263:96-100. doi: 10.1016/j.jviromet.2018.11.001. Epub 2018 Nov 2.

Abstract

Rice black-streaked dwarf virus (RBSDV) infects rice plants, a major crop, and is transmitted via the small brown planthopper (SBPH: Laodelphax striatellus Fallén), causing significant economic loss in China. To rapidly diagnose RBSDV, a reverse transcription-recombinase polymerase amplification (RT-RPA) method was developed using P10 virus-specific primers and probes. Detection of terminally labeled amplification products was achieved with the lateral flow strip method. Our results demonstrate that RT-RPA and RT-PCR assays offer similar sensitivity and specificity in RBSDV detection using cDNA as template. The optimum RT-RPA reaction temperature and time was 37 °C and 20 min, respectively. By screening twenty-one field suspected rice plants, the RT-RPA assay was confirmed to be simple, rapid and reliable. Thus, the RBSDV RT-RPA assay developed here will be a successful tool for quick diagnosis of RBSDV-infected rice plants.

Keywords: Lateral flow strip; RBSDV; Recombinase polymerase amplification.

Publication types

  • Comparative Study
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Genes, Viral / genetics
  • Molecular Diagnostic Techniques / instrumentation*
  • Molecular Diagnostic Techniques / methods*
  • Nucleic Acid Amplification Techniques*
  • Oryza / virology
  • Plant Diseases / virology*
  • Plant Viruses / genetics*
  • RNA, Viral / genetics
  • Reverse Transcriptase Polymerase Chain Reaction
  • Sensitivity and Specificity
  • Temperature
  • Time Factors

Substances

  • RNA, Viral

Supplementary concepts

  • Rice black streaked dwarf virus