Electrochemiluminescence Biosensor Based on a Duplex-Specific Nuclease and Dual-Output Toehold-Mediated Strand Displacement Cascade Amplification Strategy for Sensitive Detection of MicroRNA-499
Qian Wang, Linying Yu, Yao Peng, Mengting Sheng, Zhiying Jin, Tingting Zhang, Jianshe Huang, Xiurong Yang
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引用次数: 0
Abstract
The timely and accurate diagnosis of acute myocardial infarction (AMI) is of great significance to reduce mortality and morbidity associated with the condition. Herein, we developed an electrochemiluminescence (ECL) biosensor for the detection of the potential AMI biomarker microRNA-499 (miRNA-499), which was based on duplex-specific nuclease-assisted target recycling and dual-output toehold-mediated strand displacement (TMSD). First, miRNA-499 was converted into a large amount of single-stranded DNA through the DSN-assisted target recycling, which was further incubated with the DNA triple-stranded complex (S) to implement TMSD cycles. Thus, the Ru(bpy)32+-labeled signal strands were released and captured by the capture probe on the electrode surface, resulting in an intense ECL signal. Owing to the prominent cascade signal amplification, the constructed biosensor exhibited a good linear response to miRNA-499 within the range of 100 aM–100 pM with a detection limit of 69.99 aM. Furthermore, it demonstrated superior selectivity, stability, and reproducibility. In addition, the biosensor was successfully applied to detect miRNA-499 in real human serum samples, demonstrating its potential for nucleic acid detection in the early diagnosis of diseases.
期刊介绍:
Analytical Chemistry, a peer-reviewed research journal, focuses on disseminating new and original knowledge across all branches of analytical chemistry. Fundamental articles may explore general principles of chemical measurement science and need not directly address existing or potential analytical methodology. They can be entirely theoretical or report experimental results. Contributions may cover various phases of analytical operations, including sampling, bioanalysis, electrochemistry, mass spectrometry, microscale and nanoscale systems, environmental analysis, separations, spectroscopy, chemical reactions and selectivity, instrumentation, imaging, surface analysis, and data processing. Papers discussing known analytical methods should present a significant, original application of the method, a notable improvement, or results on an important analyte.