CRISPR-based biosensing systems: a way to rapidly diagnose COVID-19

Abstract

The outbreak of the emerging SARS-CoV-2 virus has highlighted the challenges of detecting viral infections, especially in resource-limited settings. The SARS-CoV-2 virus transmission chain is interrupted when screening and diagnosis can be performed on a large scale by identifying asymptomatic or moderately symptomatic patients. Diagnosis of COVID-19 with reverse transcription polymerase chain reaction (RT-PCR) has been limited due to inadequate access to complex, expensive equipment and reagents, which has impeded efforts to reduce the spread of virus transmission. Recently, the development of several diagnostic platforms based on the CRISPR-Cas system has reduced the dependence on RT-PCR. The first CRISPR-based diagnostic test for SARS-CoV-2 was recently approved by the U.S. Food and Drug Administration. The biosensing systems have several important features that make them suitable for point-of-care tests, including the speed of design and synthesis of each platform in less than a few days, an assay time of 1-2 h, and the cost of materials and reagents less than one dollar per test. The HUDSON-SHERLOCK and STOPCovid biosensing systems, as field-deployable and rapid diagnostic tests, can detect low-copy viruses in body fluids without nucleic acid extraction and with minimal equipment. In addition, Cas13-based treatment strategies could potentially be an effective antiviral strategy for the prevention and treatment of emerging pandemic viruses such as SARS-CoV-2. In this review, we describe recent advances in CRISPR-based diagnostic platforms with an emphasis on their use in the rapid diagnosis and potential treatment of COVID-19.

Exploring nano-enabled CRISPR-Cas-powered strategies for efficient diagnostics and treatment of infectious diseases

“…In owing to increased genotyping of SNPs, Cas14-DETEC-TR's development offers potential for utility in identifying ssDNA viruses that are therapeutically, economically, and environmentally relevant. Cas12b-based DNA detection (CD detection) has been used to directly identify plasma HPV cell less tumour DNA in Cas14-DETECTR, demonstrating that Cas12b-based DNA detection (CD detection) might assist in quickly determining viral infectiousness [168].…”

Outlook of various diagnostics and nanodiagnostic techniques for COVID-19

“…CRISPR-Cas platforms are used widely as POC technology. CRISPR systems involve a CRISPR RNA or guide RNA that binds to the target DNA or RNA sequence and effector Cas proteins with nuclease activity ( Vatankhah et al, 2020 ). Specific High-sensitivity Enzymatic Reporter UnLOCKing (SHERLOCK) assay and DNA Endonuclease-Targeted CRISPR Trans Reporter (DETECTOR) assay are the two methods developed by Sherlock Biosciences and Mammoth Biosciences, respectively, to detect viral RNA.…”

Analysis of Recent Bio-/Nanotechnologies for Coronavirus Diagnosis and Therapy

“…Several platforms have been applied for COVID-19 detection: specific high-sensitivity enzymatic reporter unlocking (SHERLOCK), HOLMES (one-hour low-cost multipurpose highly efficient system), DNA (Desoxyribonucleic acid), endonuclease-targeted CRISPR trans reporter (DETECTR), CAS-EXPAR(EXPonantial Amplification Reaction), NASBACC (nucleic acid sequences-based amplification CRISPR Cleavage), STOPCovid, ctPCR (cycle threshold PCR), and AIOD-CRISPR(All in one dual crispr). These platforms have been well discussed in a critical review by Vatankhah et al [ 52 ].…”

“…The first designed and most used platform, called SHERLOCK showed a successful application for identifying cell-free tumor DNA mutations, genotyping human DNA, detectingdengue and Zika viruses, and distinguishing pathogenic bacteria [ 52 ]. Several studies have also demonstrated the potential application of this platform for COVID-19 detection [ 53 ].…”

“…By combining SHERLOCK with the HUDSON protocol; a process for lysis of viral particles and inactivation of DNases and RNases in body fluids with heat and chemical treatment, no prior RNA extraction is needed [ 52 ]. In a recent study, HUDSON was improved to allow virus inactivation within 10 minthusproviding a simple, rapid and single step Cas-13 detection tool of COVID-19.…”