Journal
BIOSENSORS & BIOELECTRONICS
Volume 47, Issue -, Pages 324-328Publisher
ELSEVIER ADVANCED TECHNOLOGY
DOI: 10.1016/j.bios.2013.03.020
Keywords
Circular strand-displacement polymerase reaction; Hybridization chain reaction; Electrochemical DNA sensor; Signal amplification
Categories
Funding
- NSFC [21025521, 21035001, 21190041]
- National Key Basic Research Program [2011 CB911000]
- CSIRT Program
- NSF of Hunan Province [10JJ7002]
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We developed a novel electrochemical strategy for ultrasensitive DNA detection using a dual amplification strategy based on the circular strand-displacement polymerase reaction (CSDPR) and the hybridization chain reaction (HCR). In this assay, hybridization of hairpin-shaped capture DNA to target DNA resulted in a conformational change of the capture DNA with a concomitant exposure of its stem. The primer was then hybridized with the exposed stem and triggered a polymerization reaction, allowing a cyclic reaction comprising release of target DNA, hybridization of target with remaining capture DNA, polymerization initiated by the primer. Furthermore, the free part of the primer propagated a chain reaction of hybridization events between two DNA hairpin probes with biotin labels, enabling an electrochemical reading using the streptavidin-alkaline phosphatase. The proposed biosensor showed to have very high sensitivity and selectivity with a dynamic response range through 10 fM to 1 nM, and the detect limit was as low as 8 fM. The proposed strategy could have the potential for molecular diagnostics in complex biological systems. (C) 2013 Elsevier B.V. All rights reserved.
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