Three-mode electrochemical sensing of ultralow MicroRNA levels

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Journal titleJournal of the American Chemical Society
Pages30273038; # of pages: 12
SubjectCancer cells; Cancer classification; Direct detection; Dynamic range; Electrochemical sensing; High selectivity; Human serum; MicroRNAs; PCR amplification; Protein binding; Sequential analysis; Single electrodes; Wide dynamic range; Biochemistry; Electrochemical sensors; Polymerase chain reaction; RNA; Sensors; microRNA; microRNA 122; microRNA 21; microRNA 32; protein p19; unclassified drug; article; concentration (parameters); electrochemical analysis; electrochemical detection; electrode; gene amplification; human; human tissue; hybridization; limit of detection; protein binding; protein localization; real time polymerase chain reaction; RNA analysis; sensor; Base Sequence; DNA Primers; Electrochemical Techniques; Humans; MicroRNAs; Nucleic Acid Hybridization; Real-Time Polymerase Chain Reaction
AbstractMicroRNAs (miRNAs) are an emerging class of biomarkers that are frequently deregulated in cancer cells and have shown great promise for cancer classification and prognosis. In this work, we developed a three-mode electrochemical sensor for detection and quantitation of ultralow levels of miRNAs in a wide dynamic range of measured concentrations. The sensor facilitates three detection modalities based on hybridization (H-SENS), p19 protein binding (P-SENS), and protein displacement (D-SENS). The combined three-mode sensor (HPD-SENS) identifies as low as 5 aM or 90 molecules of miRNA per 30 μL of sample without PCR amplification, and can be operated within the dynamic range from 10 aM to 1 μM. The HPD sensor is made on a commercially available gold nanoparticles-modified electrode and is suitable for analyzing multiple miRNAs on a single electrode. This three-mode sensor exhibits high selectivity and specificity and was used for sequential analysis of miR-32 and miR-122 on one electrode. In addition, the H-SENS can recognize miRNAs with different A/U and G/C content and distinguish between a fully matched miRNA and a miRNA comprising either a terminal or a middle single base mutation. Furthermore, the H- and P-SENS were successfully employed for direct detection and profiling of three endogenous miRNAs, including hsa-miR-21, hsa-miR-32, and hsa-miR-122 in human serum, and the sensor results were validated by qPCR. © 2013 American Chemical Society.
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AffiliationNational Research Council Canada (NRC-CNRC); NRC Steacie Institute for Molecular Sciences (SIMS-ISSM)
Peer reviewedYes
NPARC number21269641
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Record identifier5591f322-f823-4e82-bd15-e919edd403e2
Record created2013-12-13
Record modified2016-05-09
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