期刊
BIOSENSORS & BIOELECTRONICS
卷 36, 期 1, 页码 12-17出版社
ELSEVIER ADVANCED TECHNOLOGY
DOI: 10.1016/j.bios.2012.03.032
关键词
Electrochemical biosensor; Aptasensor; Signal amplification; Primer extension; Protein; Ultrasensitivity
类别
资金
- National Basic Research Program of China [2010CB732400]
- National Natural Science Foundation of China [21075141, 21135002, 81101638, 21121091]
A simple electrochemical aptasensor was developed for ultrasensitive protein detection by combining a novel strategy of cyclic target-induced primer extension (CTIPE) with an aptamer-hairpin probe and enzyme-amplified electrochemical readout. In the presence of protein target, the immobilized aptamer-hairpin probe recognized the protein to trigger primer extension reaction by target-induced conformational transition, which released the protein from replicated DNA duplex. The released target could cyclically bind with other aptamer-hairpin probes and trigger new primer extension, leading to formation of numerous biotin-tagged DNA duplex, which significantly amplified the protein recognition event and facilitated the subsequent enzymatic signal enhancement, leading to an ultrasensitive electrochemical aptasensor. Using human vascular endothelial growth factor as a model protein, the designed aptasensor could detect protein down to 0.82 pg mL(-1) with a linear range from 1 pg mL(-1) to 1 ng mL(-1). The proposed aptasensor was amenable to quantification of protein in complex biological matrixes, and would become a simple and powerful tool for bioanalysis and clinic diagnostic application. (C) 2012 Elsevier B.V. All rights reserved.
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