Abstract
Virus disease spreads effortlessly mechanically or through minute insect vectors that are extremely challenging to avoid. Emergence and reemergence of new viruses such as severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), H1N1 influenza virus, avian influenza virus, dengue virus, Citrus tristeza virus, and Tomato yellow leaf curl virus have paralyzed the economy of many countries. The cure for major viral diseases is not feasible; however, early detection and surveillance of the disease can obstruct their spread. Therefore, advances in the field of virus diagnosis and the development of new point-of-care testing kits become necessary globally. Clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated protein (Cas) is an emerging technology for gene editing and diagnostics development. Several rapid nucleic acid diagnostic kits have been developed and validated using Cas9, Cas12, and Cas13 proteins. This review summarizes the CRISPR/Cas-based next-generation molecular diagnostic techniques and portability of devices for field-based utilization.
Highlights
For the last two decades, the emergence and reemergence of new viral diseases have compromised healthcare and agriculture worldwide
The Zika virus outbreak led Pardee et al (2016) to first use Cas9 as a diagnostic tool for viruses, where they used nucleic acid sequence-based amplification (NASBA), an isothermal amplification technique based on Clustered regularly interspaced short palindromic repeats (CRISPR) cleavage (NASBACC)
SHERLOCK (Kellner et al, 2020), DNA Endonuclease Targeted CRISPR Trans Reporter (DETECTR) (Chen et al, 2018, Broughton et al, 2020), HOLMES and HOLMESv2 (Li et al, 2018, 2019), and many other techniques commonly use fluorescent-based assays to develop POC diagnostic kits for viruses, and such methods are frequently being utilized for SARSCoV-2 diagnosis (Guo et al, 2020) (Figure 1)
Summary
For the last two decades, the emergence and reemergence of new viral diseases have compromised healthcare and agriculture worldwide. The Zika virus outbreak led Pardee et al (2016) to first use Cas9 as a diagnostic tool for viruses, where they used nucleic acid sequence-based amplification (NASBA), an isothermal amplification technique based on CRISPR cleavage (NASBACC). The establishment of inactive dead Cas9 (dCas9), which selectively binds to target DNA without nick/cleavage, enhances the popularity of this protein in the development of CRISPR/Cas9-based diagnostic kits.
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