Progress and bioapplication of CRISPR-based one-step, quantitative and multiplexed infectious disease diagnostics

Area of Science:

• Molecular Diagnostics
• Biosensing Technology
• Infectious Disease Research

Background:

• In vitro diagnosis (IVD) is crucial for early disease detection using biomarkers.
• CRISPR-Cas systems are emerging as highly sensitive and specific IVD tools for pathogen identification.
• Current research aims to enhance CRISPR-based diagnostics for practical applications.

Approach:

• Reviewing novel CRISPR-based detection strategies including extraction-free and amplification-free methods.
• Exploring modifications in Cas/crRNA complexes for improved assay performance.
• Investigating advancements in quantitative assays, one-pot detection, and multiplexed platforms.

Key Points:

• CRISPR-Cas systems offer superior sensitivity and specificity for infectious disease detection.
• Emerging approaches focus on simplifying workflows (e.g., extraction-free, amplification-free).
• Developments include quantitative assays, one-pot methods, and multiplexed platforms for comprehensive diagnostics.

Conclusions:

• Novel CRISPR-Cas approaches enhance quantification, multiplexed detection, and point-of-care testing (POCT).
• These advancements position CRISPR-Cas as next-generation diagnostic biosensing platforms.
• Future innovations will address real-world challenges, including pandemics like COVID-19.