Highly sensitive one-step DNA detection using repeated DNA amplification and Cas12a

Detection limits of 62 aM and 58 aM were achieved for the SARS-CoV-2 S and N genes, respectively.

• A modular one-pot platform integrates catalytic hairpin assembly, rolling circle amplification, and CRISPR-Cas12a for nucleic acid detection.
• The system creates a self-sustained amplification loop that enhances signal detection through real-time fluorescence.
• Single-stranded DNA probes functionalized on gold nanoparticles enable colorimetric changes and measurable signals without complex equipment.
• The platform demonstrated effective discrimination in controlled experiments, suggesting potential for point-of-care applications.
• Assay time varies from 20 to 120 minutes depending on sensitivity requirements.

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