Journal
ACS SUSTAINABLE CHEMISTRY & ENGINEERING
Volume 9, Issue 28, Pages 9257-9263Publisher
AMER CHEMICAL SOC
DOI: 10.1021/acssuschemeng.1c01454
Keywords
strand displacement amplification; ligase; miRNA; electrochemistry; nicking endonuclease
Categories
Funding
- Science and Technology Cooperation Project between the Chinese and Australian Governments [2017YFE0132300]
- Social Development Project of Jiangsu Province [BE2018629]
- Postdoctoral Program in China [2019T120461]
- Postdoctoral Program in Jiangsu Province [CRO201904]
- Top-notch Talents Program in Wuxi City [BJ2020035]
- Science and Technology Bureau of Wuxi City [WX18IIAN020]
- Program of Wuxi City Health and Family Planning Commission [2018629]
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miRNAs are powerful biomarkers for various pathological processes, with miR-200c being a critical signature of prostate cancer. An established biosensor using electrochemical techniques and ligation-dependent cascade strand displacement amplification shows high sensitivity and reliability for miRNA detection. This sensor can discriminate target miRNAs from mismatched ones and performs well in biological samples, offering a potential application for miRNA sensing.
miRNAs are emerging as powerful biomarkers, which can be used for pointing out various pathological processes of human beings. For example, miR-200c is a critical signature of prostate cancer. Nevertheless, classic approaches for an miRNA assay still suffer certain limitations. Herein, we have established a highly sensitive miR-200c biosensor utilizing electrochemical techniques. A smart sensing strategy is designed focusing on ligation-dependent cascade strand displacement amplification (CSDA). Target miR-200c is able to link two DNA probes aided by the T4 DNA ligase. The as-formed DNA hybrid is then applied for cyclic strand displacement polymerization and nicking cleavage processes in the solution. Subsequently, the generated product is able to trigger downstream SDA at the electrode interface. The combined procedure helps recruit a large number of DNA probes labeled with electrochemical species. The recorded electrochemical signal is positively related to the initial miR-200c level. Under optimal conditions, the as-prepared biosensor achieves a high sensitivity. The limit of detection (LOD) is estimated to be as low as 3.3 aM. Moreover, it successfully discriminates the target miRNA from mismatched miRNAs and performs satisfactorily in biological samples. The working electrode can be easily regenerated for additional tests by heating and annealing processes. This work offers a facile, reliable, and highly sensitive way to sense miRNAs for potential applications.
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