Graphene oxide assisted light-up aptamer selection against Thioflavin T for label-free detection of microRNA

Sci Rep. 2021 Feb 22;11(1):4291. doi: 10.1038/s41598-021-83640-z.

Abstract

We selected an aptamer against a fluorogenic dye called Thioflavin T (ThT). Aptamers are single-stranded DNA that can bind a specific target. We selected the ThT aptamer using graphene oxide assisted SELEX and a low-cost Open qPCR instrument. We optimized, minimized, and characterized the best aptamer candidate against ThT. The aptamer, ThT dye, and the enzymatic strand displacement amplification (SDA) were used in a label-free approach to detect the micro RNA miR-215 in saliva and serum. The aptamer confers higher specificity than intercalating dyes but without expensive covalently modified DNA probes. This isothermal, low-cost, simple method can detect both DNA and RNA. The target, miR-215, was detected with a limit of detection of 2.6 nM.

Publication types

  • Research Support, N.I.H., Extramural

MeSH terms

  • Aptamers, Nucleotide* / chemistry
  • Base Sequence
  • Benzothiazoles* / chemistry
  • Biosensing Techniques*
  • DNA / analysis
  • DNA Probes
  • Fluorescent Dyes
  • Graphite / chemistry*
  • MicroRNAs / analysis
  • MicroRNAs / chemistry*
  • Nanotechnology / methods
  • Nanotechnology / standards
  • Sensitivity and Specificity
  • Spectrum Analysis

Substances

  • Aptamers, Nucleotide
  • Benzothiazoles
  • DNA Probes
  • Fluorescent Dyes
  • MicroRNAs
  • graphene oxide
  • thioflavin T
  • Graphite
  • DNA