G-quadruplex-based assay combined with aptamer and gold nanoparticles for Escherichia coli K88 determination

Mikrochim Acta. 2020 May 1;187(5):308. doi: 10.1007/s00604-020-04291-x.

Abstract

A colorimetric method was developed using G-quadruplex and gold nanoparticles (AuNPs) for determination of Escherichia coli K88 (ETEC K88). It was composed of two modules: (1) an aptamer as biorecognizing element and (2) a capturing DNA (modified with AuNPs at 5') as a transducer. In the absence of target bacteria, the aptamer can form stable double strands with capturing DNA, preventing the binding of capturing DNA to the G-quadruplex. However, the double strands of capturing DNA and aptamer are untied due to the stronger binding of aptamers to bacteria in the presence of target bacteria. As a result, the G-quadruplex binds to capture DNA and leads to the aggregation and color change of AuNPs, which can be monitored by a spectrophotometer or visualization. The quantitative determination was achieved by monitoring the optical density change of AuNPs solution at 524 nm after target addition. Under optimal conditions, the method has a low detection limit (1.35 × 102 CFU mL-1) and a linear response in the range 102 to 106 CFU mL-1. Graphical abstract The manuscripts describe a colorimetric method for the detection of ETEC K88 by using intermolecular G-quadruplex to induce the agglomeration of gold nanoparticles, which can be directly used to determine the presence of bacteria with our naked eyes.

Keywords: Aptamer; Bacteria; Colorimetric method; Intermolecular G-quadruplex; Visualization.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Aptamers, Nucleotide / chemistry*
  • Biosensing Techniques / methods
  • Colorimetry / methods*
  • DNA / chemistry*
  • Escherichia coli / isolation & purification*
  • G-Quadruplexes*
  • Gold / chemistry
  • Humans
  • Limit of Detection
  • Metal Nanoparticles / chemistry*
  • Ponds / microbiology
  • Spectrophotometry / methods
  • Urine / microbiology
  • Water Pollutants / analysis

Substances

  • Aptamers, Nucleotide
  • Water Pollutants
  • Gold
  • DNA