Visual detection of norovirus genogroup II by reverse transcription loop-mediated isothermal amplification with hydroxynaphthol blue dye

Food Environ Virol. 2014 Sep;6(3):196-201. doi: 10.1007/s12560-014-9142-8. Epub 2014 Apr 22.

Abstract

A simple, rapid, specific, and sensitive colorimetric reverse transcription loop-mediated isothermal amplification (RT-LAMP) assay with hydroxynaphthol blue dye (HNB) was established, targeting RNA-dependent RNA polymerase and capsid protein gene for the detection of the dominant norovirus genogroup in China-NoV GII. The assay was carried out at 65 °C for 60 min with no cross-reactivity with other common gastroenteritis viruses. The sensitivity of this assay was 10(3) copies per reaction which is equivalent to the conventional RT-PCR test. The clinical test showed 94.83% coincidence rate for NoV genogroup II detection compared with the results, confirmed by the Department of Viral Diarrhea of Chinese Center for Disease Control and Prevention via conventional RT-PCR. The HNB dye-based RT-LAMP could be a novel rapid screening method for prevalent norovirus genogroup II in China, especially in those resource-limited hospitals and rural local clinics.

Publication types

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

MeSH terms

  • Caliciviridae Infections / virology*
  • China
  • Coloring Agents / chemistry
  • Coloring Agents / metabolism*
  • Genotype
  • Humans
  • Naphthalenesulfonates / chemistry
  • Naphthalenesulfonates / metabolism*
  • Norovirus / chemistry
  • Norovirus / genetics
  • Norovirus / isolation & purification*
  • Nucleic Acid Amplification Techniques / instrumentation
  • Nucleic Acid Amplification Techniques / methods*
  • Reverse Transcription
  • Viral Proteins / genetics

Substances

  • Coloring Agents
  • Naphthalenesulfonates
  • Viral Proteins
  • trisodium 3-hydroxy-4-((2Z)-2-(2-oxo-4-sulfonatonaphthalen-1-ylidene)hydrazinyl)naphthalene-2,7-disulfonate