CRISPR/Cas12-Driven Exponential Amplification Combined with a Lateral Flow Biosensor Enabling Rapid and Highly...

Area of Science:

• Biotechnology
• Molecular Diagnostics
• Biosensor Technology

Background:

• Accurate DNA detection is crucial for research and clinical diagnostics.
• CRISPR technology enhances DNA testing speed, portability, and accuracy.
• Existing methods may lack sensitivity or require complex equipment.

Purpose of the Study:

• To develop a rapid and highly sensitive DNA detection biosensor.
• To leverage Cas12 activity and exponential amplification for enhanced detection.
• To create a point-of-care (POC) compatible diagnostic tool.

Main Methods:

• Developed a Cas12-driven exponential amplification-based lateral flow biosensor (CADEX-LF).
• Utilized Cas12's target-loading-dependent *trans*-cleavage activity.
• Employed nicking endonuclease-mediated exponential amplification for signal enhancement.

Main Results:

• Achieved a detection sensitivity of 2 × 10^-15 M within 45 minutes.
• Demonstrated outstanding specificity with double-base resolution.
• Successfully identified herpes simplex virus 1 (HSV-1) DNA in clinical samples.

Conclusions:

• CADEX-LF provides rapid, sensitive, and specific DNA detection.
• The biosensor is suitable for point-of-care applications, requiring no complex equipment.
• CADEX-LF shows significant potential for clinical diagnosis, particularly in resource-limited environments.