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Abstract:
A new signal amplification strategy based on target-regulated DNA proximity hybridization (TRPH) reaction accompanying formation of three-way DNA junction was designed for electronic detection of Microcystin-LR (MC-LR used in this case), coupling with junction-induced isothermal cycling signal amplification. Initially, a sandwiched-type immunoreaction was carried out in a low-cost PCR tube between anti-MC-LR mAb(1) antibody-labeled DNA(1) (mAb(1)-DNA(1)) and anti-MC-LR mAb(2)-labeled DNA(2) (mAb(2)-DNA(2)) in the presence of target to form a three-way DNA junction. Then, the junction could undergo an unbiased strand displacement reaction on an h-like DNA nanostructure-modified electrode (labeled with methylene blue redox tag on the short DNA strand), thereby resulting in the dissociation of methylene blue-labeled signal DNA from the electrode. The newly formed double-stranded DNA could be cleaved again by exonuclease III, and the released three-way DNA junction retriggered the strand-displacement reaction with h-like DNA nanostructures for junction recycling. During the strand-displacement reaction, numerous methylene blue-labeled DNA strands were far away from the electrode, thus decreasing the detectable electrochemical signal within the applied potentials. Under optimal conditions, the TRPH-based immunosensing system exhibited good electrochemical responses for detecting target MC-LR at a concentration as low as 1.0 ng kg(-1) (1.0 ppt). Additionally, the precision, reproducibility, specificity and method accuracy were also investigated with acceptable results. (C) 2015 Elsevier B.V. All rights reserved.
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BIOSENSORS & BIOELECTRONICS
ISSN: 0956-5663
Year: 2015
Volume: 69
Page: 241-248
7 . 4 7 6
JCR@2015
1 0 . 7 0 0
JCR@2023
ESI Discipline: CHEMISTRY;
ESI HC Threshold:265
JCR Journal Grade:1
CAS Journal Grade:1
Cited Count:
WoS CC Cited Count: 32
SCOPUS Cited Count:
ESI Highly Cited Papers on the List: 0 Unfold All
WanFang Cited Count:
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30 Days PV: 0
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