期刊
SCIENTIFIC REPORTS
卷 6, 期 -, 页码 -出版社
NATURE PUBLISHING GROUP
DOI: 10.1038/srep35982
关键词
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资金
- National Natural Science Foundation of China [21535002, 21605071, 21545004]
- Special Funds for the Construction of Taishan Scholars [tspd20150209]
- Shandong Provincial Natural Science Foundation [ZR2014BL025]
- Opening fund of Shandong Provincial Key Laboratory of Detection Technology for Tumor Markers [KLDTTM2015-8]
MicroRNA (miRNA) was a promising class of cancer biomarkers. Here we developed a label-free method for sensitive measurement of let-7d miRNA based on multiple amplification techniques. The primer will bind to the duplex strand DNA that was formed by stem-loop template and target let-7d to initiate strand displacement amplification (SDA) in tandem. The released single strand DNA will be a primer to bind the circular template to initiate rolling circle amplification (RCA). The products based on multiple amplifications will be detected by a standard fluorimeter with N-methyl mesoporphyrin IX (NMM) as the fluorescent indicator. The proposed method exhibited excellent selectivity and high sensitivity with a detection limit of as low as 1.5 x 10(-13) M. Moreover, this methodology was used for the determination of biomolecules in real serum samples with satisfying results.
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