Three-dimensional DNA nanomachine biosensor coupled with CRISPR Cas12a cascade amplification for ultrasensitive detection of carcinoembryonic antigen

J Nanobiotechnology. 2024 May 18;22(1):266. doi: 10.1186/s12951-024-02535-z.

Abstract

The detection of carcinoembryonic antigen (CEA) holds significant importance in the early diagnosis of cancer. However, current methods are hindered by limited accessibility and specificity. This study proposes a rapid and convenient Cas12a-based assay for the direct detection of CEA in clinical serum samples, aiming to address these limitations. The protocol involves a rolling machine operation, followed by a 5-min Cas12a-mediated cleavage process. The assay demonstrates the capability to detect human serum with high anti-interference performance and a detection limit as low as 0.2 ng/mL. The entire testing procedure can be accomplished in 75 min without centrifugation steps, and successfully reduced the limit of detection of traditional DNA walking machine by 50 folds. Overall, the testing procedure can be easily implemented in clinical settings.

Keywords: CEA; CRISPR-Cas12a; Cascade amplification; DNA rolling machine.

MeSH terms

  • Bacterial Proteins / genetics
  • Biosensing Techniques* / methods
  • CRISPR-Associated Proteins
  • CRISPR-Cas Systems*
  • Carcinoembryonic Antigen* / blood
  • DNA* / chemistry
  • Endodeoxyribonucleases
  • Humans
  • Limit of Detection*
  • Nucleic Acid Amplification Techniques / methods

Substances

  • Cas12a protein