Programmable no-nonspecific genetic analytical system via dual-circle-based rolling circle amplification with an efficient CRISPR/Cas12a biosensing strategy

Nov 19, 2025Scientific reports

A precise genetic analysis method using two linked DNA circles and a sensitive CRISPR-based detection system

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Abstract

The achieved a low detection limit of 0.77 fM for DNA.

  • The system integrates nicking endonuclease-mediated amplification with CRISPR/Cas12a for enhanced specificity.
  • It demonstrates a wide linear range of detection from 10 fM to 1 nM.
  • The method specifically recognizes single mismatched DNA, reducing the risk of false positives.
  • In serum samples, results align well with those from real-time quantitative polymerase chain reaction (qPCR).
  • This approach could be advantageous for cancer screening in settings with limited resources.

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Key numbers

10 fM to 1 nM
Detection Range
Wide linear detection range for gene analysis.
0.77 fM
Limit of Detection
Lowest detection limit achieved in the study.

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