1. REVEALR: A Multicomponent XNAzyme-Based Nucleic Acid Detection System for SARS-CoV-2
- Author
-
John C. Chaput and Kefan Yang
- Subjects
Male ,Paper ,Analyte ,Loop-mediated isothermal amplification ,Computational biology ,010402 general chemistry ,Sensitivity and Specificity ,01 natural sciences ,Biochemistry ,Catalysis ,chemistry.chemical_compound ,Colloid and Surface Chemistry ,Limit of Detection ,Nasopharynx ,Chlorocebus aethiops ,Animals ,Humans ,CRISPR ,Vero Cells ,Detection limit ,SARS-CoV-2 ,COVID-19 ,RNA ,DNA, Catalytic ,General Chemistry ,Nucleic acid amplification technique ,0104 chemical sciences ,Oligodeoxyribonucleotides ,chemistry ,COVID-19 Nucleic Acid Testing ,Nucleic acid ,RNA, Viral ,Female ,Nucleic Acid Amplification Techniques ,DNA - 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.
- Published
- 2021