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Construction of a Novel Biosensor Based on the Self-assembly of Dual-Enzyme Cascade Amplification-Induced Copper Nanoparticles for Ultrasensitive Detection of MicroRNA153.

Abstract
MicroRNAs (miRNAs) have received extensive attention because of their potential as biomarkers for cancer diagnosis and monitoring, and their effective detection is very significant. Here, a specific, one-pot, rapid, femtomolar sensitive miRNAs detection biosensor was developed based on the target-triggered three-way junction (3-WJ) and terminal deoxynucleotide transferase (TDT)/Nt.BspQI in combination with activated copper nanoparticles (CuNPs) self-assembly. To this end, a 3-WJ hairpin probe and helper probe were designed to selectively identify the target miRNA, so as to form a stable 3-WJ structure that further triggered the double-enzyme cycling to produce poly T to activate the self-assembly of CuNPs. Based on the simplicity of CuNPs generation, the poly T template fluorescence CuNPs can detect the minimum detection limit of 1 fm within 1.75 h. In addition, the applicability of this method in complex samples was demonstrated by analyzing the whole-blood RNA extraction from Parkinson patients, consisting of the results of commercial miRNA kits. The developed strategy performs powerful implications for miRNA detection, which may be beneficial for the effective diagnostic assays and biological research of Parkinson's disease.
AuthorsJingyu Cui, Houyu Han, Jiafang Piao, Hai Shi, Dianming Zhou, Xiaoqun Gong, Jin Chang
JournalACS applied materials & interfaces (ACS Appl Mater Interfaces) Vol. 12 Issue 30 Pg. 34130-34136 (Jul 29 2020) ISSN: 1944-8252 [Electronic] United States
PMID32627523 (Publication Type: Journal Article)
Chemical References
  • Biomarkers
  • DNA Probes
  • Enzymes, Immobilized
  • MIRN153 microRNA, human
  • MicroRNAs
  • Poly T
  • Copper
  • DNA Nucleotidylexotransferase
  • DNA Restriction Enzymes
Topics
  • Biomarkers (blood)
  • Biosensing Techniques (methods)
  • Copper (chemistry)
  • DNA Nucleotidylexotransferase (chemistry, metabolism)
  • DNA Probes (metabolism)
  • DNA Restriction Enzymes (metabolism)
  • Enzymes, Immobilized (chemistry, metabolism)
  • Humans
  • Limit of Detection
  • Metal Nanoparticles (chemistry)
  • MicroRNAs (analysis, blood)
  • Parkinson Disease (diagnosis, genetics)
  • Poly T (chemistry)
  • Spectrometry, Fluorescence

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