One-step fabrication of trimetallic alloy nanozyme catalyzer for luminol-H2O2 chemiluminescence and its application for miRNA-21 detection coupled with miRNA walking machine

J Pharm Biomed Anal. 2020 Jul 15:186:113280. doi: 10.1016/j.jpba.2020.113280. Epub 2020 Mar 26.

Abstract

PtCuCo trimetallic alloys (PtCuCo-TAs) are synthesized by one-step reduction. The chemiluminescence (CL) properties of PtCuCo-TAs are studied systemically. PtCuCo-TAs show good catalyzing for luminol-H2O2 system. A CL platform is developed for the detection of miRNA-21 using PtCuCo-TAs as nanozyme catalyzer. In the CL detection platform, H1 (Hairpin DNA1) is immobilized onto magnetic beads (MBs) firstly. In the presence of miRNA-21, H1 is opened. H2 (Hairpin DNA2) then hybridizes with H1. Meanwhile, a "cleat" in the end of miRNA-21 with a fewer bases complementary is formed to prevent miRNA-21 dissociating from H1. This miRNA-21 hybridizes to another H1. When cpDNA-PtCuCo-TAs which consisted with cDNA (Complementary strand of probe DNA) and pDNA-PtCuCo-TAs (PtCuCo-TAs labeled with probe DNA) are added, the ssDNA region of H1 reacts with the toehold domain of probe DNA and cDNA is released resulting pDNA-PtCuCo-TAs being captured. With this process repeatedly, a lot of pDNA-PtCuCo-TAs are captured onto MBs. After separation and washing, the precipitate and H2O2 are put into the 96-well and luminol solution is injected. The CL signal is produced by PtCuCo-TAs catalyzing luminol-H2O2 system. The amount of miRNA-21 is detected with CL signal. This CL platform performs with limit of detection 0.167 fM and has good selectivity over other RNA.

Keywords: Enzyme-free; Nanocatalyst; One-step; Trimetallic alloys; miRNA-21.

MeSH terms

  • Alloys / chemistry
  • DNA / chemistry*
  • Humans
  • Hydrogen Peroxide / chemistry
  • Limit of Detection
  • Luminescence
  • Luminescent Measurements / methods*
  • Luminol / chemistry
  • Metal Nanoparticles*
  • MicroRNAs / analysis*

Substances

  • Alloys
  • MIRN21 microRNA, human
  • MicroRNAs
  • Luminol
  • DNA
  • Hydrogen Peroxide