Colorimetric detection of norovirus by helicase-dependent amplification method based on specific primers integrated with HRPzyme

Anal Bioanal Chem. 2022 Sep;414(23):6723-6733. doi: 10.1007/s00216-022-04247-5. Epub 2022 Aug 5.

Abstract

Noroviruses (NoVs) are the most common causes of epidemic gastroenteritis, responsible for at least 50% of all gastroenteritis outbreaks worldwide and significant causes of foodborne illness. In the USA, approximately 21 million illnesses attributable to NoVs have annually occurred. Therefore, there is a great demand to develop a rapid, low-cost, and accurate detection method for NoVs. This study first reported colorimetric helicase-dependent amplification (HDA) methods based on specific primers integrated with HRPzyme for the rapid and sensitive detection of NoV GI and GII. The colorimetric HDA methods exhibited a detection limit of 10 copies mL-1 of each NoV GI and GII and were confirmed to be specific to each NoV GI and GII. The period required to complete the HDA method was 2 h, including a step of RNA extraction and cDNA synthesis without expensive instruments such as a thermal cycler and detector. The cutoff value of the method for the oyster artificially inoculated with a known amount of NoV was all 102 copies g-1 for NoV GI and GII. Therefore, the HDA method developed in this study can be useful tool for the on-site detection of NoVs in food samples.

Keywords: Colorimetric detection; HRPzyme; Helicase-dependent amplification; Norovirus (NoV).

MeSH terms

  • Caliciviridae Infections* / diagnosis
  • Caliciviridae Infections* / epidemiology
  • Colorimetry
  • DNA Primers / genetics
  • Gastroenteritis* / epidemiology
  • Genotype
  • Humans
  • Norovirus* / genetics
  • Phylogeny
  • RNA, Viral / genetics

Substances

  • DNA Primers
  • RNA, Viral