Rapid and sensitive detection of COVID-19 using CRISPR/Cas12a-based detection with naked eye readout, CRISPR/Cas12a-NER

CRISPR-Cas System: An Approach With Potentials for COVID-19 Diagnosis and Therapeutics

“…Cas13 could process the CRISPR array to yield individual crRNA for targeting multiple viral RNAs. There have been reports on the induction of mutations in target sites by CRISPR-based antivirals using Cas9 (Wang et al, 2018;Wang et al, 2020), while no crRNA target-site mutation was observed following Cas13 treatment (Freije et al, 2019). These reports indicate the potential of the Cas13 enzyme as an effective antiviral to manage SARS-CoV-2 infection.…”

Next-generation pathogen diagnosis with CRISPR/Cas-based detection methods

“…For example, integrating later flow assay (CORDS, CRISPR/Cas12a-LFD) and naked-eye detection (CRISPR/Cas-based colorimetric platform, fluorescence-based POC system), four groups quickly developed Cas12a-based on-site diagnostic assays for African swine virus (ASFV), an emergence virus for the global swine industry (Table 2) [44][45][46][47]. For the current COVID-19 pandemic [48], several research groups are developing CRISPR/Cas-based diagnostic assay [49][50][51][52][53]. Among which, these diagnostic methods can detect as low as 2 copies of extracted SARS-CoV-2 RNA and readout on later flow strip in an hour without requiring for elaborate instrumentation or visualized by green fluorescent with naked-eye under 485 nm light [49][50][51][52].…”

“…For the current COVID-19 pandemic [48], several research groups are developing CRISPR/Cas-based diagnostic assay [49][50][51][52][53]. Among which, these diagnostic methods can detect as low as 2 copies of extracted SARS-CoV-2 RNA and readout on later flow strip in an hour without requiring for elaborate instrumentation or visualized by green fluorescent with naked-eye under 485 nm light [49][50][51][52]. More recently, Joung and colleagues developed an integrated test called STOPCovid (SHER-LOCK Testing in One Pot) for detecting SARS-CoV-2, which can return result on the strip in 70 min, with a detection limit of 100 copies of viral genome input in saliva or nasopharyngeal swabs per reaction [53].…”

Molecular Diagnosis of COVID-19: An Update and Review

“…To enable onsite diagnosis, an ssDNA reporter labeled with a quenched green fluorescent molecule is used which is cleaved by Cas12a; when there is nucleic acid of SARS-CoV-2, it results in green fluorescence under 485 nm light. 33,34…”