Construction of electrochemical aptasensor of carcinoembryonic antigen based on toehold-aided DNA recycling signal amplification

Bioelectrochemistry. 2020 Jun:133:107492. doi: 10.1016/j.bioelechem.2020.107492. Epub 2020 Feb 21.

Abstract

Carcinoembryonic antigen (CEA), serves as a broad-spectrum tumor marker, and plays an important role in reflecting the existence, therapeutic evaluation, development, monitoring and prognosis of many types of cancer. An electrochemical aptasensor was designed for CEA detection based on toehold-aided DNA recycling. A partially hybridized Probe-4 (i.e. P2/P3/P4) was self-assembled on the surface of a gold electrode serving as the sensing platform. For CEA detection, CEA can bind with aptamer and free probe-1 (P1) can hybridize with P4, triggering toehold-aided DNA recycling. This enables the hybridization of more probe-5 (P5) (labeled with methylene blue (MB)) with P4, causing more methylene blue (MB) to be brought close to the electrode surface. An amplified current signal was thus generated due to more MB in the electrode surface. The proposed design showed good linearity between current response and log CEA concentration ranging from 0.1 to 50 ng·mL-1, with a detection limit of 20 pg mL-1. This aptasensor also showed high specificity for CEA detection, and was successfully used in spiked biological samples.

Keywords: Cancer; Carcinoembryonic antigen; Electrochemical aptasensor; Methylene blue; Strand displacement reaction.

MeSH terms

  • Aptamers, Nucleotide / chemistry*
  • Biosensing Techniques / methods*
  • Carcinoembryonic Antigen / analysis*
  • Carcinoembryonic Antigen / blood
  • Carcinoembryonic Antigen / urine
  • Electrochemical Techniques / methods
  • Humans
  • Limit of Detection
  • Nucleic Acid Hybridization / methods
  • Saliva / chemistry

Substances

  • Aptamers, Nucleotide
  • Carcinoembryonic Antigen