Fluorescence biosensor to detect microRNAs via integrating DNA hairpins transition mediated strand displacement amplification with primer exchange reaction

Bioorg Med Chem Lett. 2024 Jul 1:106:129774. doi: 10.1016/j.bmcl.2024.129774. Epub 2024 Apr 28.

Abstract

Herein, we constructed a fluorescence biosensor for the ultra-sensitive analysis of microRNAs (miRNAs) by combining DNA hairpins transition triggered strand displacement amplification (DHT-SDA) with primer exchange reaction (PER). Target miRNA initiated DHT-SDA to facilitate the generation of multiple single-stranded DNA (ssDNA) as PER primer, which was extended into a long ssDNA. The biosensor is successfully utilized in detecting miRNAs with high sensitivity (limit of detection for miRNA-21 was 58 fM) and a good linear relationship between 100 nM and 100 fM. By simply changing the DNA hairpin sequence, the constructed biosensor can be extended to analyze another miRNAs. Moreover, the biosensor has the feasibility of detecting miRNAs in real samples with satisfactory accuracy and reliability. Therefore, the fluorescent biosensor has great application potential in clinical diagnosis.

Keywords: DNA hairpins transition; Detection; Primer exchange reaction; Strand displacement amplification; miRNA-21.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Biosensing Techniques*
  • DNA / chemistry
  • DNA Primers / chemistry
  • DNA, Single-Stranded / chemistry
  • DNA, Single-Stranded / metabolism
  • Fluorescence
  • Humans
  • Inverted Repeat Sequences
  • Limit of Detection
  • MicroRNAs* / analysis
  • MicroRNAs* / metabolism
  • Nucleic Acid Amplification Techniques*
  • Spectrometry, Fluorescence

Substances

  • MicroRNAs
  • DNA
  • DNA, Single-Stranded
  • DNA Primers