期刊
NANOTECHNOLOGY
卷 27, 期 42, 页码 -出版社
IOP PUBLISHING LTD
DOI: 10.1088/0957-4484/27/42/425502
关键词
isothermal amplification; TAIEA; miRNA detection; fluorescent biosensor; CuNPs
资金
- Mid-career Researcher Support Program through the National Research Foundation (NRF) - Ministry of Science, ICT and Future Planning (MSIP) of Korea [2015R1A2A1A01005393]
- Bio-Synergy Research Project through the NRF - MISP of Korea [NRF-2015M3A9C4070484]
- Industrial Source Technology Development Program of the Ministry of Trade, Industry and Energy (MOTIE) [2010-10038662]
- Korea Evaluation Institute of Industrial Technology (KEIT) [10038662] Funding Source: Korea Institute of Science & Technology Information (KISTI), National Science & Technology Information Service (NTIS)
- National Research Foundation of Korea [2015R1A2A1A01005393] Funding Source: Korea Institute of Science & Technology Information (KISTI), National Science & Technology Information Service (NTIS)
We devised a novel method for rapid and ultrasensitive detection of target microRNA (miRNA) by employing target-assisted isothermal exponential amplification (TAIEA) combined with poly (thymine)-templated fluorescent copper nanoparticles (CuNPs) as signaling probes. The target miRNA hybridizes to the unimolecular template DNA and works as a primer for the extension reaction to form double-stranded product, which consequently generates two nicking endonuclease recognition sites. By simultaneous nicking and displacement reactions, exponential amplification generates many poly (thymine) strands as final products, which are employed for the synthesis of fluorescent CuNPs. Based on the fluorescent signal from CuNPs, target miRNA is detected as low as 0.27 fM around 1 h of total analysis time. The diagnostic capability of this system has been successfully demonstrated by reliably detecting target miRNA from different cell lysates, showing its great potential towards real clinical applications.
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