Localized Dual-Cycle Amplification Integrating Entropy-Driven Reaction and Catalytic Hairpin Assembly for miRNA Imaging in Living Cells and Tissues

Anal Chem. 2026 May 12;98(18):13296-13306. doi: 10.1021/acs.analchem.5c06796. Epub 2026 Apr 27.

Abstract

Intracellular microRNA (miRNA) detection plays a crucial role in cancer diagnosis and treatment monitoring. However, current methods face challenges in sensitivity, specificity, and cellular application. Here, we present a novel localized dual-cycle amplification (LDA) system integrating localized entropy-driven DNA circuit reaction (LEDR) and localized catalytic hairpin assembly (LCHA) for sensitive miRNA imaging in living cells. This approach employs a dual-nanosphere platform functionalized with EDR and CHA units. The presence of target miR-155 initiates a cascade reaction of LEDR-LCHA process. The LDA system demonstrates several advantages: (1) enhanced sensitivity with a detection limit of 1.42 pM for miR-155; (2) enhanced reaction kinetics due to localized strategy, reaching signal plateau within 15 min under in vitro conditions; (3) efficient cellular uptake without transfection reagents and improved stability in biological fluids. The system successfully monitored intracellular miR-155 levels and discriminated between cancer and normal samples in both living cells and clinical tissues. This LDA system achieves a synergistic integration of sensitivity, rapid response, and practical applicability for intracellular miRNA detection, offering a robust platform for cancer diagnostics.

MeSH terms

  • Catalysis
  • Entropy*
  • Humans
  • MicroRNAs* / analysis
  • MicroRNAs* / genetics
  • Nucleic Acid Amplification Techniques* / methods

Substances

  • MicroRNAs
  • MIRN155 microRNA, human