A novel quantification platform for point-of-care testing of circulating MicroRNAs based on allosteric spherical nanoprobe

J Nanobiotechnology. 2020 Oct 31;18(1):158. doi: 10.1186/s12951-020-00717-z.

Abstract

MiRNA-150, a gene regulator that has been revealed to be abnormal expression in non-small cell lung cancer (NSCLC), can be regarded as a serum indicator for diagnosis and monitoring of NSCLC. Herein, a new sort of nanoprobe, termed allosteric spherical nanoprobe, was first developed to sense miRNA-150. Compared with conventional hairpin, this new nanoprobe possesses more enrichment capacity and reaction cross section. Structurally, it consists of magnetic nanoparticles and dual-hairpin. In the absence of miRNA-150, the spherical nanoprobes form hairpin structure through DNA self-assembly, which could promote the Förster resonance energy transfer (FRET) of fluorophore (FAM) and quencher (BHQ1) nearby. However, in the presence of target, the target-probe hybridization can open the hairpin and form the active "Y" structure which separated fluorophore and quencher to yield "signal on" fluorescence. In the manner of multipoint fluorescence detection, the target-bound allosteric spherical nanoprobe could provide high detection sensitivity with a linear range of 100 fM to 10 nM and a detection limit of 38 fM. More importantly, the proposed method can distinguish the expression of serum miRNA-150 among NSCLC patients and healthy people. Finally, we hoped that the potential bioanalytical application of this nanoprobe strategy will pave the way for point-of-care testing (POCT).

Keywords: Allosteric spherical nanoprobe; Dual-hairpin; Förster resonance energy transfer (FRET); MiRNA-150; Point-of-care testing (POCT).

MeSH terms

  • Allosteric Site
  • Biomarkers, Tumor / blood*
  • Biosensing Techniques
  • Carcinoma, Non-Small-Cell Lung / diagnosis*
  • Circulating MicroRNA / blood*
  • Female
  • Fluorescence Resonance Energy Transfer
  • Fluorescent Dyes / chemistry*
  • Humans
  • In Situ Hybridization, Fluorescence
  • Limit of Detection
  • Lung Neoplasms / diagnosis*
  • Magnetic Iron Oxide Nanoparticles / chemistry*
  • Male
  • MicroRNAs / blood*
  • Middle Aged
  • Point-of-Care Testing
  • Surface Properties

Substances

  • Biomarkers, Tumor
  • Circulating MicroRNA
  • Fluorescent Dyes
  • MIRN150 microRNA, human
  • MicroRNAs