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Please use this identifier to cite or link to this item: http://ntour.ntou.edu.tw:8080/ir/handle/987654321/38519

Title: Using Self-Assembled Aptamers and Fibrinogen-Conjugated Gold Nanoparticles to Detect DNA Based on Controlled Thrombin Activity
Authors: Chuan-Kuo Chen
Yen-Chun Shiang
Chih-Ching Huang
Huan-Tsung Chang
Contributors: 國立臺灣海洋大學:生命科學暨生物科技學系
Keywords: Single-nucleotide polymorphism
Gold nanoparticles
Enzyme
Self-assembly
DNA sensor
Date: 2011
Issue Date: 2016-09-08T06:36:33Z
Publisher: Biosensors and Bioelectronics
Abstract: Abstract: We have developed a colorimetric probe, based on the aggregation of gold nanoparticles (Au NPs), for the detection of DNA and for the analysis of single-nucleotide polymorphism (SNP); this probe functions through the modulation of the activity of thrombin (Thr) in the presence of bivalent thrombin-binding aptamers (TBAs). The bivalent TBAs were formed from TBA27′ (comprising a 27-base sequence providing TBA27 functionality, a T5 linker, and an 11-base sequence for hybridization) and TBA15′ (comprising a 15-base sequence providing TBA15 functionality, a T5 linker, and a 12-base sequence for hybridization) through their hybridization with perfectly matched DNA (DNApm). The bivalent TBAs interacted specifically with thrombin, suppressing its activity toward fibrinogen-modified Au NPs (Fib-Au NPs). The potency of the inhibitory effect of TBA15′–TBA27′/DNApm toward thrombin – and, thus, the degree of aggregation of the Fib-Au NPs – was highly dependent on the concentration of DNApm. Under the optimal conditions (50 pM thrombin, 2 nM TBA15′, 2 nM TBA27′, and 38 pM Fib-Au NPs), the linear relationship of the response of the probe toward DNApm extended from 0.1 to 2 nM, with a correlation coefficient of 0.97. The limit of detection (LOD) for DNApm was 20 pM, based on a signal-to-noise ratio of 3. We also applied a corresponding TBA15″–TBA27″/Thr/Fib-Au NP probe to the detection of the SNP of the Arg249Ser unit in the TP53 gene, with an LOD of 32 pM. Relative to conventional molecular beacon-based and crosslinking aggregation-based Au NP probes, our new approach offers higher sensitivity and higher selectivity toward DNA.
Relation: 26(8)
URI: http://ntour.ntou.edu.tw:8080/ir/handle/987654321/38519
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