title: Ultrasensitive and Multiple Disease-Related MicroRNA Detection Based on Tetrahedral DNA Nanostructures and Duplex-Specific Nuclease-Assisted Signal Amplification.
abstract: A highly sensitive and multiple microRNA (miRNA) detection method by combining three-dimensional (3D) DNA tetrahedron-structured probes (TSPs) to increase the probe reactivity and accessibility with duplex-specific nuclease (DSN) for signal amplification for sensitive miRNA detection was proposed. Briefly, 3D DNA TSPs labeled with different fluorescent dyes for specific target miRNA recognition were modified on a gold nanoparticle (GNP) surface to increase the reactivity and accessibility. Upon hybridization with a specific target, the TSPs immobilized on the GNP surface hybridized with the corresponding target miRNA to form DNA-RNA heteroduplexes, and the DSN can recognize the formed DNA-RNA heteroduplexes to hydrolyze the DNA in the heteroduplexes to produce a specific fluorescent signal corresponding to a specific miRNA, while the released target miRNA strands can initiate another cycle, resulting in a significant signal amplification for sensitive miRNA detection. Different targets can produce different fluorescent signals, leading to the development of a sensitive detection for multiple miRNAs in a homogeneous solution. Under optimized conditions, the proposed assay can simultaneously detect three different miRNAs in a homogeneous solution with a logarithmic linear range spanning 5 magnitudes (10-12-10-16) and achieving a limit of detection down to attomolar concentrations. Meanwhile, the proposed miRNA assay exhibited the capability of discriminating single bases (three bases mismatched miRNAs) and showed good eligibility in the analysis of miRNAs extracted from cell lysates and miRNAs in cell incubation media, which indicates its potential use in biomedical research and clinical analysis.
authors.affiliation: Research Center for Bioengineering and Sensing Technology, School of Chemistry and Bioengineering, University of Science & Technology Beijing , Beijing 100083, P. R. China.
authors.forename: Fang
authors.initials: F
authors.lastname: Xu
authors.affiliation: Research Center for Bioengineering and Sensing Technology, School of Chemistry and Bioengineering, University of Science & Technology Beijing , Beijing 100083, P. R. China.
authors.forename: Haifeng
authors.initials: H
authors.lastname: Dong
authors.affiliation: Research Center for Bioengineering and Sensing Technology, School of Chemistry and Bioengineering, University of Science & Technology Beijing , Beijing 100083, P. R. China.
authors.forename: Yu
authors.initials: Y
authors.lastname: Cao
authors.affiliation: School of Space and Environment, Beihang University , Beijing 100191, P. R. China.
authors.forename: Huiting
authors.initials: H
authors.lastname: Lu
authors.affiliation: Research Center for Bioengineering and Sensing Technology, School of Chemistry and Bioengineering, University of Science & Technology Beijing , Beijing 100083, P. R. China.
authors.forename: Xiangdan
authors.initials: X
authors.lastname: Meng
authors.affiliation: Research Center for Bioengineering and Sensing Technology, School of Chemistry and Bioengineering, University of Science & Technology Beijing , Beijing 100083, P. R. China.
authors.forename: Wenhao
authors.initials: W
authors.lastname: Dai
authors.affiliation: Research Center for Bioengineering and Sensing Technology, School of Chemistry and Bioengineering, University of Science & Technology Beijing , Beijing 100083, P. R. China.
authors.forename: Xueji
authors.initials: X
authors.lastname: Zhang
authors.affiliation: Department of Zoology, College of Science, P.O. Box 2455, King Saud University , Riyadh 11451, Saudi Arabia.
authors.forename: Khalid Abdullah
authors.initials: KA
authors.lastname: Al-Ghanim
authors.affiliation: Department of Zoology, Government College University , Faisalabad, Pakistan.
authors.forename: Shahid
authors.initials: S
authors.lastname: Mahboob
mesh_terms: D004247:DNA; D006046:Gold; D053768:Metal Nanoparticles; D035683:MicroRNAs; D049329:Nanostructures; D021141:Nucleic Acid Amplification Techniques
chemical_list: D035683:MicroRNAs; D006046:Gold; D004247:DNA
keywords: duplex-specific nuclease; multiple microRNA detection; signal amplification; tetrahedral DNA nanostructures; ultrasensitive microRNA detection