Competitive aptamer bioassay for selective detection of adenosine triphosphate based on metal-paired molecular conformational switch and fluorescent gold nanoclusters

Biosensors & Bioelectronics
Jing-Min Liu, Xiu-Ping Yan

Abstract

A competitive aptamer bioassay was developed for the selective detection of adenosine triphosphate (ATP). The proposed bioassay employed the T-Hg-T induced hairpin-structure as the molecule conformational switch (MCS), aptamer as a specific recognizer, and mercaptoundecanoic acid modified gold nanoclusters (MUA-AuNCs) as a sensitive signal reporter. The T-rich MCS ssDNA with the sequence complementary with that for the aptamer of ATP was bound with Hg(2+) to form the metal-paired hairpin-structure. Addition of the aptamer and its target biomolecule ATP resulted in a competitive aptamer bioassay. The aptamer competed with Hg(2+) to hybridize with T-rich MCS ssDNA, thereby destroyed the hairpin-structure. As a result, the Hg(2+) was released and the signal transduction was achieved. The ATP affected the interaction between aptamer and hairpin-structure, thus mediated the release of Hg(2+), which was sensitively quantified by fluorescent MUA-AuNCs. Under selected conditions, the developed method allowed sensitive and selective detection of ATP with a linear range of 100-2000 nM and a detection limit (3s) of 48 nM. The relative standard deviation for sixty replicate detections of 200 nM ATP was 2.1%, and the recoveries of the spike...Continue Reading

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Jan 1, 2013·Biosensors & Bioelectronics·Li-Yi ChenHuan-Tsung Chang
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