CRISPR/Cas13a combined with hybridization chain reaction for visual detection of influenza A (H1N1) virus

Anal Bioanal Chem. 2022 Dec;414(29-30):8437-8445. doi: 10.1007/s00216-022-04380-1. Epub 2022 Oct 20.

Abstract

This study provides proof of concept of a colorimetric biosensor for influenza H1N1 virus assay based on the CRISPR/Cas13a system and hybridization chain reaction (HCR). Target RNA of influenza H1N1 virus activated the trans-cleavage activity of Cas13a, which cleaved the special RNA sequence (-UUU-) of the probe, further initiating HCR to copiously generate G-rich DNA. Abundant G-quadruplex/hemin was formed in the presence of hemin, thus catalyzing a colorimetric reaction. The colorimetric biosensor exhibited a linear relationship from 10 pM to 100 nM. The detection limit was 0.152 pM. The biosensor specificity was excellent. This new and sensitive detection method for influenza virus is a promising rapid influenza diagnostic test.

Keywords: CRISPR/Cas13a; DNAzyme; Hybridization chain reaction; Influenza H1N1 virus detection.

MeSH terms

  • Biosensing Techniques* / methods
  • Clustered Regularly Interspaced Short Palindromic Repeats
  • DNA, Catalytic* / metabolism
  • G-Quadruplexes*
  • Hemin
  • Humans
  • Influenza A Virus, H1N1 Subtype* / genetics
  • Influenza, Human* / diagnosis

Substances

  • Hemin
  • DNA, Catalytic