期刊
TALANTA
卷 235, 期 -, 页码 -出版社
ELSEVIER
DOI: 10.1016/j.talanta.2021.122728
关键词
MicroRNA detection; SERS; Duplex-specific nuclease signal amplification; Iodide-modified Ag nanoparticles
资金
- National Natural Science Foundation of China [21874025, 21775028]
- Science and Technol-ogy Commission of Shanghai Municipality [17JC1401900]
- Nature Science Foundation of Zhejiang Province [LQ20B050002]
A new detection method for miRNA-21 was proposed in this study, achieving ultra-sensitive and quantitative analysis through DSN signal amplification strategy and SERS technology. The method showed excellent performance in detecting miRNA-21 and was successfully applied for monitoring the expression levels of miRNA-21 in different cancer cell lines and human serum.
With the emergence of microRNA (miRNA) as a key player in early clinical disease diagnosis, development of rapidly sensitive and quantitative miRNA detection methods are imperative. Herein, a label-free SERS assay coupled with duplex-specific nuclease (DSN) signal amplification strategy was proposed for facilely ultrasensitive and quantitative analysis of miRNA-21. Firstly, magnetic beads assembled with excessive capture DNA were utilized to hybridize the target miRNA-21. These DNA-RNA heteroduplexes were cleaved by DSN to generate small nucleotide fragments into the supernatant and the miRNA-21 released and rehybridized another DNA, going to the next DSN cycle. Consequently, numerous of small nucleotide fragments of capture DNA were released from magnetic beads and the miRNA-21 signal was transferred and amplified by the SERS signals of total phosphate backbones which are abundant in nucleotide. Furthermore, iodide-modified Ag nanoparticles (AgINPs) was employed to generate a strong and reproducible SERS signal. The proposed method displayed excellent performance for miRNA-21 detection with the linear range from 0.33 fM to 3.3 pM, and a lower detection limit of 42 aM. Moreover, this strategy exhibited effectively base discrimination capability and was successfully applied for monitoring the expression levels of miRNA-21 in different cancer cell lines and human serum.
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