Development and application of sensitive, specific, and rapid CRISPR-Cas13-based diagnosis

Area of Science:

• Biotechnology
• Molecular Diagnostics
• Genomics

Background:

• Current nucleic acid detection methods are inadequate for timely diagnosis of infectious diseases like COVID-19 and cancer mutations.
• The COVID-19 pandemic highlighted the need for accessible and rapid diagnostic tools.
• Slow and costly mutation detection hinders personalized cancer treatment.

Purpose of the Study:

• To review the development and applications of CRISPR-Cas13 assays for medical diagnosis.
• To highlight the advantages of CRISPR-Cas13-based diagnostics over traditional methods.
• To present real-world examples of CRISPR-Cas13 assay performance.

Main Methods:

• CRISPR-Cas13 system utilizing crRNA-guided RNA targeting and collateral cleavage activity.
• Detection of target RNA via collateral cleavage of a fluorescent reporter.
• Adaptation of lateral flow strips for portability and micro-well chips for high-throughput multiplexing.

Main Results:

• CRISPR-Cas13 assays demonstrate >95% sensitivity and >99% specificity.
• Rapid detection (<2 hours) at a low cost ($0.05 per test).
• Assays are portable and adaptable for high-throughput applications.

Conclusions:

• CRISPR-Cas13 assays represent a significant advancement in nucleic acid detection for medical and fieldwork.
• This technology offers a superior alternative to RT-PCR for various diagnostic applications.
• CRISPR-Cas13 holds promise for improved infectious disease surveillance and personalized medicine.