Journal
ANALYTICA CHIMICA ACTA
Volume 878, Issue -, Pages 95-101Publisher
ELSEVIER SCIENCE BV
DOI: 10.1016/j.aca.2015.04.018
Keywords
MicroRNAs; Electrochemical biosensors; Single enzyme amplification; Redox cycling; Alkaline phosphatase
Categories
Funding
- National Natural Science Foundation of China [21205003, 21305004]
- Joint Fund for Fostering Talents of National Natural Science Foundation of China [U1304205]
- Education Department of Henan Province [14A150042]
- Program for Science and Technology Innovation Talents at the University of Henan Province [15HASTIT001]
- Joint Fund for Fostering Talents of National Natural Science Foundation of Henan Province [U1304205]
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MicroRNAs (MiRNAs) have been regarded as clinically important biomarkers and drug discovery targets. In this work, we reported a simple and ultrasensitive electrochemical method for miRNAs detection based on single enzyme amplification and electrochemical-chemical-chemical (ECC) redox cycling. Specifically, upon contact with the target miRNAs, the hairpin structure of biotinylated DNA immobilized on gold electrode was destroyed and the biotin group in DNA was forced away from the electrode surface, allowing for the coupling of streptavidin-conjugated alkaline phosphatase (SA-ALP). Then, ascorbic acid (AA, the enzymatic product of ALP) triggered the ECC redox cycling with ferrocene methanol (FcM) and tris(2-carboxyethyl) phosphine (TCEP) as the redox mediator and the chemical reducing reagent, respectively. The method was more sensitive than that with horseradish peroxidase (HRP) or glucose oxidase (GOx) triggered recycling since one ALP molecule captured by one target miRNA molecule promoted the production of thousands of AA. Analytical merits (e.g., detection limit, dynamic range, specificity, regeneration and reproducibility) were evaluated. The feasibility of the method for analysis of miRNA-21 in human serum has also been demonstrated. (C) 2015 Elsevier B.V. All rights reserved.
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