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Sensors 2015, 15(2), 2629-2643; doi:10.3390/s150202629

A Novel Ultrasensitive ECL Sensor for DNA Detection Based on Nicking Endonuclease-Assisted Target Recycling Amplification, Rolling Circle Amplification and Hemin/G-Quadruplex

Department of Clinical Laboratory, Xinqiao Hospital, Third Military Medical University, Chongqing 400037, China
These authors contributed equally to this work.
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Author to whom correspondence should be addressed.
Received: 1 December 2014 / Accepted: 19 January 2015 / Published: 26 January 2015
(This article belongs to the Special Issue Biosensors for Pathogen Detection)
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Abstract

In this study, we describe a novel universal and highly sensitive strategy for the electrochemiluminescent (ECL) detection of sequence specific DNA at the aM level based on Nt.BbvCI (a nicking endonuclease)-assisted target recycling amplification (TRA), rolling circle amplification (RCA) and hemin/G-quadruplex. The target DNAs can hybridize with self-assembled capture probes and assistant probes to form “Y” junction structures on the electrode surface, thus triggering the execution of a TRA reaction with the aid of Nt.BbvCI. Then, the RCA reaction and the addition of hemin result in the production of numerous hemin/G-quadruplex, which consume the dissolved oxygen in the detection buffer and result in a significant ECL quenching effect toward the O2/S2O82− system. The proposed strategy combines the amplification ability of TRA, RCA and the inherent high sensitivity of the ECL technique, thus enabling low aM (3.8 aM) detection for sequence-specific DNA and a wide linear range from 10.0 aM to 1.0 pM. At the same time, this novel strategy shows high selectivity against single-base mismatch sequences, which makes our novel universal and highly sensitive method a powerful addition to specific DNA sequence detection. View Full-Text
Keywords: target recycling amplification (TRA); rolling circle amplification (RCA); hemin/G-quadruplex; Nt.BbvCI; ECL target recycling amplification (TRA); rolling circle amplification (RCA); hemin/G-quadruplex; Nt.BbvCI; ECL
This is an open access article distributed under the Creative Commons Attribution License which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. (CC BY 4.0).

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MDPI and ACS Style

Luo, F.; Xiang, G.; Pu, X.; Yu, J.; Chen, M.; Chen, G. A Novel Ultrasensitive ECL Sensor for DNA Detection Based on Nicking Endonuclease-Assisted Target Recycling Amplification, Rolling Circle Amplification and Hemin/G-Quadruplex. Sensors 2015, 15, 2629-2643.

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