Development of a homogeneous time-resolved fluorescence assay for detection of viral double-stranded RNA

Anal Biochem. 2019 Feb 1:566:46-49. doi: 10.1016/j.ab.2018.10.021. Epub 2018 Oct 21.

Abstract

The group of positive-sense single-stranded RNA ((+) ssRNA) viruses includes many important human pathogens. However, specific antiviral agents are not currently available for many RNA viruses. For screening of antiviral agents, methods that are simple, rapid, and compatible with high-throughput are required. Here, we describe a novel method for measurement of double-stranded RNA using a homogeneous time-resolved fluorescence assay. This method allowed detection of human rhinovirus (HRV), enterovirus, coxsackievirus, and murine norovirus. Furthermore, this method detected antiviral activity of a HRV 3C protease inhibitor. The assay may be useful for discovery of antiviral agents against (+) ssRNA viruses.

Keywords: Antiviral agents; Double-stranded RNA; Homogeneous time-resolved fluorescence assay; RNA virus; Viral replication.

MeSH terms

  • 3C Viral Proteases
  • Antiviral Agents / chemistry
  • Cysteine Endopeptidases
  • Fluorescence
  • Fluoroimmunoassay / methods*
  • Protease Inhibitors / chemistry
  • RNA Viruses / isolation & purification*
  • RNA, Double-Stranded / analysis*
  • RNA, Viral / analysis*
  • Viral Proteins / antagonists & inhibitors

Substances

  • Antiviral Agents
  • Protease Inhibitors
  • RNA, Double-Stranded
  • RNA, Viral
  • Viral Proteins
  • Cysteine Endopeptidases
  • 3C Viral Proteases