期刊
BIOSENSORS & BIOELECTRONICS
卷 102, 期 -, 页码 211-216出版社
ELSEVIER ADVANCED TECHNOLOGY
DOI: 10.1016/j.bios.2017.11.030
关键词
Ratiometric; Electrochemical assay; Dual-amplification; microRNA
类别
资金
- National Natural Science Foundation of China [21365015, 21475056, 21775065]
- Program for Major Academic and Technical Leaders of Jiangxi Province [20123BCB22003, 20162BCB22013]
- Key Project of Scientific and Technological Innovation Talents in Jiangxi Province [20165BCB18022]
- Landing Project of Science and Technology of Colleges in Jiangxi Province [KJLD13010, KJLD14009]
We propose a ratiometric electrochemical assay for detecting microRNA (miRNA) on the basis of dual-amplification mechanism by using distinguishable electrochemical signals from thionine (Thi) and ferrocene (Fc). The thiol-modified and ferrocene-labeled hairpin capture probes (CP) are first immobilized on an Au electrode via Au-S reaction. The target miRNA hybridizes with CP and unfolding the hairpin structure of CP to form miRNA-DNA duplexes. Then, kamchatka crab duplex specific nuclease (DSN) specifically cleaves the DNA in miRNA-DNA duplexes, leading to the release of miRNA and another cleaves cycle, meanwhile, numerous Fc leaves away from the electrode surface and leads to the signal-off of Fc. The residual fragment on electrode surface acts as a HCR primer to form dsDNA polymers through in situ HCR with the presence of the primer and two probes (HDNA and HDNA'), resulting in the capture of numerous DNA/Au NPs/Thi and the signal-on of Thi. The dual amplification mechanism significantly amplifies the decrease of Fc signal and the increase of Thi signal for ratiometric readout (I-Thi/I-Fc), thus providing a sensitive method for the selective detection of miR-141 with a detection limit down to 11 aM. The dual-signal ratiometric outputs have an intrinsic self-calibration to the effects from system, which is promising to be applied in biosensing and clinical diagnosis.
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