Yu Han, Xuefeng Bian, Xiao Wang, Jiaying Chen, Liu Han, Xin Sun
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引用次数: 0
Abstract
Due to the limiting factors associated with microRNAs (miRNAs), including their low abundance, short sequence length, and high family sequence homology, the gold standard for miRNA detection does not meet the requirements for rapid and low-cost clinical detection of miRNAs in biological samples. To this end, we developed a one-pot method called molecular aptamer beacon-utilized exponential isothermal amplification (MABEXIA) for analyzing and detecting miRNAs. This novel devolved method was analyzed using both a fluorescent dye and a horseradish peroxidase (HRP) mediated chemiluminescence reaction with luminol and hydrogen peroxide for miRNA detection. The principle of the fluorescence platform involves hybridization of the target miRNA with MAB, which results in the formation of a large amount of double-stranded (ds) DNA products under the dual catalytic action of a DNA polymerase (Klenow Fragment) and nicking endonuclease (Nb.BbvCI) at a constant temperature. Moreover, this method exhibited good detection performance and was successfully applied to 1% human serum for miRNA detection. In contrast to the fluorescence method, the chemiluminescence approach achieved miRNA detection through the targeted interaction between the activated streptavidin (SA) aptamer within the dsDNA product and SA-coated magnetic beads. This method showed a good linear relationship in the ranges of 50 fM to 50 pM for miRNA detection. Finally, this chemiluminescence approach has a detection ability comparable to that of the gold standard for miRNA detection.
期刊介绍:
Organic & Biomolecular Chemistry is an international journal using integrated research in chemistry-organic chemistry. Founded in 2003 by the Royal Society of Chemistry, the journal is published in Semimonthly issues and has been indexed by SCIE, a leading international database. The journal focuses on the key research and cutting-edge progress in the field of chemistry-organic chemistry, publishes and reports the research results in this field in a timely manner, and is committed to becoming a window and platform for rapid academic exchanges among peers in this field. The journal's impact factor in 2023 is 2.9, and its CiteScore is 5.5.