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Sensors 2018, 18(7), 2179; https://doi.org/10.3390/s18072179

A Non-Label and Enzyme-Free Sensitive Detection Method for Thrombin Based on Simulation-Assisted DNA Assembly

1
School of Computer Science, Shaanxi Normal University, Xi’an 710119, China
2
College of Life Sciences, Shaanxi Normal University, Xi’an 710119, China
3
National Engineering Laboratory for Resource Developing of Endangered Chinese Crude Drugs in Northwest of china, Shaanxi Normal University, Xi’an 710119, China
*
Author to whom correspondence should be addressed.
Received: 20 May 2018 / Revised: 30 June 2018 / Accepted: 3 July 2018 / Published: 6 July 2018
(This article belongs to the Special Issue Label-Free Biosensors)
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Abstract

Taking advantage of the high selectivity of aptamers and enzyme-free catalyzed hairpin assembly (CHA) amplification strategy, we herein describe a label-free and enzyme-free sensitive fluorescent and colorimetric strategy for thrombin detection in this paper. In the presence of target, the corresponding aptamer of the partial dsDNA probes will bind to the target and liberate the initiation strand, which is artfully designed as the “on” switch for hairpin assembly. Moreover, the displaced initiation strand partakes in a multi-cycle process and produces numerous G-quadruplexes, which have a remarkable enhancement in fluorescent/colorimetric signal from NMM (N-methyl-mesoporphyrin IX) and TMB (3,3′,5,5′-tetramethylbenzidine), respectively. The proposed amplification strategy for thrombin detection is of high sensitivity, down to 2.4 pM, and also achieves colorimetric signals that are able to be distinguished by naked eye. More importantly, the thermodynamics of interacting DNA strands used in our work, and the process of toehold strand displacement-driven assembly are simulated before biological testing, verifying the feasibility theoretically, and simplifying the subsequent actual experiments. Therefore, our approach and simulation have a certain potential application in biomarker detection and quantitatively monitor for disease diagnosis. View Full-Text
Keywords: thrombin detection; fluorescence; colorimetric; isothermal cycling signal amplification; computation and simulation thrombin detection; fluorescence; colorimetric; isothermal cycling signal amplification; computation and simulation
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Zhang, Y.; Wang, L.; Wang, Y.; Dong, Y. A Non-Label and Enzyme-Free Sensitive Detection Method for Thrombin Based on Simulation-Assisted DNA Assembly. Sensors 2018, 18, 2179.

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