REVEALR: A Multicomponent XNAzyme-Based Nucleic Acid Detection System for SARS-CoV-2

J Am Chem Soc. 2021 Jun 23;143(24):8957-8961. doi: 10.1021/jacs.1c02664. Epub 2021 Jun 10.

Abstract

Isothermal amplification strategies capable of rapid, inexpensive, and accurate nucleic acid detection provide new options for large-scale pathogen detection, disease diagnosis, and genotyping. Here we report a highly sensitive multicomponent XNA-based nucleic acid detection platform that combines analyte preamplification with X10-23-mediated catalysis to detect the viral pathogen responsible for COVID-19. The platform, termed RNA-Encoded Viral Nucleic Acid Analyte Reporter (REVEALR), functions with a detection limit of ≤20 aM (∼10 copies/μL) using conventional fluorescence and paper-based lateral flow readout modalities. With a total assay time of 1 h, REVEALR provides a convenient nucleic acid alternative to equivalent CRISPR-based approaches, which have become popular methods for SARS-CoV-2 detection. The assay shows no cross-reactivity for other in vitro transcribed respiratory viral RNAs and functions with perfect accuracy against COVID-19 patient-derived clinical samples.

Publication types

  • Research Support, N.I.H., Extramural

MeSH terms

  • Animals
  • COVID-19 / diagnosis*
  • COVID-19 Nucleic Acid Testing / instrumentation
  • COVID-19 Nucleic Acid Testing / methods*
  • Chlorocebus aethiops
  • DNA, Catalytic / chemistry*
  • Female
  • Humans
  • Limit of Detection
  • Male
  • Nasopharynx / virology
  • Nucleic Acid Amplification Techniques
  • Oligodeoxyribonucleotides / chemistry
  • Paper
  • RNA, Viral / analysis*
  • SARS-CoV-2 / chemistry*
  • Sensitivity and Specificity
  • Vero Cells

Substances

  • DNA, Catalytic
  • Oligodeoxyribonucleotides
  • RNA, Viral