Nuclease-assisted target recycling signal amplification strategy for graphene quantum dot-based fluorescent detection of marine biotoxins

Mikrochim Acta. 2021 Mar 9;188(4):118. doi: 10.1007/s00604-020-04684-y.

Abstract

Saxitoxin (STX) is a major marine toxin from shellfish, and it is responsible for paralytic shellfish poisoning (PSP). In this study, a highly sensitive and rapid aptamer assay was developed for STX detection by combining fluorescence resonance energy transfer (FRET) and nuclease-assisted target recycling signal amplification. The aptamer STX-41 conjugated with graphene quantum dots (GQDs) was adsorbed on magnetic reduced graphene oxide (MRGO) to establish a fluorescence quenching system. Then, the binding between STX and aptamer induced the desorption of GQD-aptamer from MRGO and the restoring of fluorescence for the fluorescent determination of STX. The digestion of the target bound aptamer by DNase I could release the target for recycling thus achieving signal amplification. Under the optimized conditions, the aptamer assay showed a wide detection range (0.1-100 ng·mL-1), low detection limit (LOD of 0.035 ng·mL-1), high specificity, good recovery (86.75-94.08% in STX-spiked clam samples) and repeatability (RSD of 4.27-7.34%). Combined with fluorescent detection technology, signal amplification technology, and magnetic separation technology, the proposed method can be used to detect STX in seafood products successfully.

Keywords: Aptamer; DNase I-catalyzed target recycling signal amplification; Fluorescence resonance energy transfer; Graphene quantum dot; Saxitoxin.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Aptamers, Nucleotide / chemistry
  • Biosensing Techniques / methods*
  • Bivalvia / chemistry
  • Endodeoxyribonucleases / chemistry
  • Fluorescence Resonance Energy Transfer / methods
  • Fluorescent Dyes / chemistry*
  • Food Contamination / analysis
  • Graphite / chemistry*
  • Limit of Detection
  • Magnetic Phenomena
  • Quantum Dots / chemistry*
  • Saxitoxin / analysis*
  • Saxitoxin / chemistry

Substances

  • Aptamers, Nucleotide
  • Fluorescent Dyes
  • graphene oxide
  • Saxitoxin
  • Graphite
  • Endodeoxyribonucleases