A simple and rapid method to assay SARS-CoV-2 RNA based on a primer exchange reaction

Abstract

A simple method is proposed in this work for the detection of SARS-CoV-2 RNA based on primer exchange reaction (PER). By ingeniously integrating the PER cascade and CRISPR/cas12a system, this...

Nanotechnology-based diagnostic methods for coronavirus: From nucleic acid extraction to amplification

“…AuNPs-based lateral flow assays (LFAs) can be also used to detect SARS-CoV-2 with naked-eye readout. Li et al developed a microfluidic platform based on RT-RPA, CRISPR-Cas12a, and LFA for the contamination-free and visual detection of the SARS-CoV-2 genome( Li et al, 2022 ). In this method, the viral RNA was first extracted from swab samples.…”

“…(A) A detection approach based on the CRISPR-Cas12a system for naked-eye readout of COVID-19( Wang et al, 2020a , Wang et al, 2020b ) (Copyright, 2020 Elsevier, reproduced with permission from Elsevier Ltd.). (B) Principle of the PER-based assay for the detection of SARS-CoV-2 RNA ( Li et al, 2022 ) (Copyright, 2022 Royal Society of Chemistry, reproduced with permission from Royal Society of Chemistry Ltd.). (C) Overview of the digital warm-start-CRISPR assay( Ding et al, 2021 ) (Copyright, 2021 Elsevier, reproduced with permission from Elsevier Ltd.).…”

“…Li et al developed a technique for the detection of SARS-CoV-2 RNA by combining PER with CRISPER-Cas12a ( Fig. 8 B)( Li et al, 2022 ). When the target RNA was present, the PER cascade reaction was triggered to produce a significant amount of ssDNAs.…”

Construction of a dual-functional dumbbell probe-based fluorescent biosensor for cascade amplification detection of miRNAs in lung cancer cells and tissues

“…14 Unlike polymerase/exonuclease/nickase (PEN) reaction networks that require sophisticated coordination among multiple enzymes and complex design of probes to achieve the cyclic generation of a specific sequence, the PER strategy can realize the cyclic synthesis of any sequence with the assistance of a hairpin, a short primer (7–9 nt) and a strand displacing polymerase, providing a robust platform for engineering molecular DNA circuits. 15 The powerful PER has been widely applied for biosensing of SARSCoV-2, 16 exosomal miRNA, 17 and survivin mRNA, 15 but the detection sensitivity is limited by its linear amplification nature. As revolutionary gene-editing tools, the RNA-guided CRISPR–Cas systems have shown great promise for accurate nucleic acid diagnosis.…”

A functional RNA/DNA circuit for one-pot detection of SARS-CoV-2 RNA

“…3 In order to develop simple and rapid detection methods, a series of new nucleic acid detection technologies have emerged, 4,5 such as reverse transcription recombinase polymerase amplification (RT-RPA), 6 reverse transcription loop-mediated isothermal amplification (RT-LAMP) and CRISPR/Cas-based technology. 7,8 Although these technologies are faster and more sensitive, there are still some drawbacks. For example, RT-RPA and RT-LAMP usually require the participation of multiple enzymes, complex primer design and the inevitable reverse transcription process, which increase the cost and complexity of the detection.…”