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

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 (similar to 10 copies/mu L) 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.

Automated summary

The study introduces a highly sensitive multicomponent XNA-based nucleic acid detection platform, termed REVEALR, with a detection limit of <= 20 aM and a total assay time of 1 h for detecting the viral pathogen responsible for COVID-19. This platform offers a convenient nucleic acid alternative to equivalent CRISPR-based approaches and shows perfect accuracy against COVID-19 patient-derived clinical samples with no cross-reactivity to other in vitro transcribed respiratory viral RNAs.