An insertion approach electrochemical aptasensor for mucin 1 detection based on exonuclease-assisted target recycling

被引:67
作者
Wen, Wei
Hu, Rong
Bao, Ting
Zhang, Xiuhua
Wang, Shengfu [1 ]
机构
[1] Hubei Univ, Hubei Collaborat Innovat Ctr Adv Organ Chem Mat, Minist Educ, Key Lab Synth & Applicat Organ Funct Mol, Wuhan 430062, Peoples R China
基金
中国国家自然科学基金;
关键词
Electrochemical aptasensor; Insertion approach; Exonuclease; Target recycling; Mucin; 1; SINGLE-STRANDED-DNA; TUMOR-MARKER; SIGNAL AMPLIFICATION; ULTRASENSITIVE DETECTION; ENZYMATIC AMPLIFICATION; MODIFIED ELECTRODE; QUANTUM DOTS; CANCER-CELLS; BIOSENSOR; APTAMERS;
D O I
10.1016/j.bios.2015.04.001
中图分类号
Q6 [生物物理学];
学科分类号
071011 ;
摘要
In this work, a sensitive exonuclease-assisted amplification electrochemical aptasensor through insertion approach was developed for the detection of mucin 1 (MUC 1). In order to construct the aptasensor, 6-Mercapto-1-hexanol (MCH) was used to block partial sites of gold electrode (GE), followed by thiolated capture probe self-assembled on GE. Methylene blue (MB) labeled aptamer hybridized with capture probe at both ends to form double-strand DNA. For the MB labeled termini was close to GE, the electrochemical response was remarkable. The presence of MUC 1 caused the dissociation of the double-strand DNA owing to the specific recognition of aptamer to MUC 1. Then exonuclease I (Exo I) selectively digested the aptamer which bound with MUC 1, the released MUC 1 participated new binding with the rest aptamer. Insertion approach improved the reproducibility and Exo I-catalyzed target recycling improved the sensitivity of the aptasensor significantly. Under optimal experimental conditions, the proposed aptasensor had a good linear correlation ranged from 10 pM to 1 mu M with a detection limit of 4 pM (Signal to Noise ratio, S/N=3). The strategy had great potential for the simple and sensitive detection of other cancer markers. (C) 2015 Elsevier B.V. All rights reserved.
引用
收藏
页码:13 / 17
页数:5
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