Towards deployable CRISPR-based nucleic acid detection.

Area of Science:

• Biotechnology
• Molecular Diagnostics
• Infectious Disease Control

Background:

• Deployable diagnostics are crucial for infectious disease management, as highlighted by the COVID-19 pandemic.
• Current nucleic acid diagnostics, while sensitive, require laboratory infrastructure and trained staff, limiting their use in resource-limited areas.
• CRISPR-based diagnostics present a promising solution for accessible, rapid, and accurate nucleic acid testing.

Purpose of the Study:

• To review recent advances in CRISPR-based diagnostics for deployable applications.
• To highlight innovations addressing limitations in resource-limited settings.
• To discuss future opportunities for improving CRISPR diagnostic accessibility.

Main Methods:

• Review of innovations in sample processing to reduce equipment needs.
• Exploration of one-pot isothermal and amplification-free CRISPR detection methods.
• Analysis of multiplexing strategies for simultaneous pathogen detection.
• Consideration of reagent lyophilization and alternative readout technologies.

Main Results:

• Advances in sample processing simplify testing across diverse sample types and pathogens.
• One-pot and amplification-free approaches offer trade-offs for different deployment scenarios.
• Multiplexing enables simultaneous detection of multiple infectious agents.
• Lyophilization and novel readouts enhance deployability by minimizing cold chain requirements and enabling field use.

Conclusions:

• CRISPR-based diagnostics are advancing rapidly toward practical, deployable solutions for infectious disease testing.
• Key innovations focus on simplifying workflows, enhancing stability, and enabling versatile detection methods.
• Further development is needed to overcome remaining challenges and fully realize the potential of CRISPR diagnostics in global health settings.