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Proceedings Paper

Direct detection of microRNAs using isothermal amplification and molecular beacon with excellent sensitivity and specificity
Author(s): Wancun Zhang; Qi Zhang; Zhiyu Qian; Yueqing Gu
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Paper Abstract

MicroRNAs (miRNAs) play important roles in a wide range of biological processes, including proliferation, development, metabolism, immunological response, tumorigenesis, and viral infection. The detection of miRNAs is imperative for gaining a better understanding of the functions of these biomolecules and has great potential for the early diagnosis of human disease as well as the discovery of new drugs through the use of miRNAs as targets. In this article, we develop a highly sensitive, and specific miRNA assay based on the two-stage isothermal amplification reactions and molecular beacon. The two-stage isothermal amplification reactions involves two templates and two-stage amplification reactions under isothermal conditions. The first template enables the amplification of miRNA, and the second template enables the conversion of miRNA to the reporter oligonucleotide(Y). Importantly, different miRNAs can be converted to the same Y seperately, which can hybridize with the same set of molecular beacon to generate fluorescent signals. This assay is highly sensitive and specific with a detection limit of 1 fM and can even discriminate single-nucleotide differences. Moreover, in combination with the specific templates, this method can be applied for multiplex miRNA assay by simply using the same molecular beacon. This method has potential to become a promising miRNA quantification method in biomedical research and clinical diagnosis.

Paper Details

Date Published: 21 February 2017
PDF: 10 pages
Proc. SPIE 10079, Reporters, Markers, Dyes, Nanoparticles, and Molecular Probes for Biomedical Applications IX, 100790U (21 February 2017); doi: 10.1117/12.2251311
Show Author Affiliations
Wancun Zhang, China Pharmaceutical Univ. (China)
Qi Zhang, China Pharmaceutical Univ. (China)
Zhiyu Qian, Nanjing Univ. of Aeronautics and Astronautics (China)
Yueqing Gu, China Pharmaceutical Univ. (China)


Published in SPIE Proceedings Vol. 10079:
Reporters, Markers, Dyes, Nanoparticles, and Molecular Probes for Biomedical Applications IX
Samuel Achilefu; Ramesh Raghavachari, Editor(s)

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