Journal
ANALYTICAL CHEMISTRY
Volume 92, Issue 7, Pages 5302-5310Publisher
AMER CHEMICAL SOC
DOI: 10.1021/acs.analchem.9b05849
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Funding
- National Natural Science Foundation of China [21375063, 21575069]
- Natural Science Foundation of Jiangsu Province [BK20181382]
- China Scholarship Council (CSC) [201906865009]
- Research Fund of Jiangsu Key Laboratory for Biosensors
- Jiangsu Collaborative Innovation Center of Biomedical Functional Materials
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MicroRNAs (miRNAs) in cancer cell-derived exosomes are important cancer biomarkers. Herein, a sensitive hybridization chain reaction (HCR) electrochemical assay was fabricated for the detection of exosomal microRNA-122 (miR-122). The hairpin DNA (hpDNA) probes were first immobilized on the surface of a gold electrode. In the presence of miR-122, the hairpin structure of the hpDNA could be opened and triggered the HCR through the cross-opening and hybridization of two helper DNA hairpins. Long nicked double helixes generated from HCR are used to capture more RuHex and increase the signal of differential pulse voltammetry (DPV). In this assay, the density of the hpDNA probes on the surface of the gold electrode was precisely controlled by the simultaneous immobilization of hpDNA and short 12 nucleotides single-stranded DNA (S-12), providing a very high amplification efficiency. More importantly, the false positive signal could be reduced or completely eliminated by applying exonuclease I (Exo I) before the introduction of target miR-122. Under optimal conditions, the assay offers very high sensitivity with an attomolar level detection limit, a linear range with 9 orders of magnitude, and specificity in single mismatch discrimination. This sensitive electrochemical assay could successfully evaluate the miR-122 concentration in different cancerderived exosomes, indicating its potential use in cancer diagnostics.
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