title: A novel ultrasensitive ECL sensor for DNA detection based on nicking endonuclease-assisted target recycling amplification, rolling circle amplification and hemin/G-quadruplex.
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/S2O8(2-) 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.
authors: Department of Clinical Laboratory, Xinqiao Hospital, Third Military Medical University, Chongqing 400037, China. luofukang@tmmu.edu.cn.
authors: Fukang
authors: F
authors: Luo
authors: Department of Clinical Laboratory, Xinqiao Hospital, Third Military Medical University, Chongqing 400037, China. guimin_xiang@sina.cn.
authors: Guimin
authors: G
authors: Xiang
authors: Department of Clinical Laboratory, Xinqiao Hospital, Third Military Medical University, Chongqing 400037, China. xiaoyun_pu@sina.cn.
authors: Xiaoyun
authors: X
authors: Pu
authors: Department of Clinical Laboratory, Xinqiao Hospital, Third Military Medical University, Chongqing 400037, China. yxyxyx_668@163.com.
authors: Juanchun
authors: J
authors: Yu
authors: Department of Clinical Laboratory, Xinqiao Hospital, Third Military Medical University, Chongqing 400037, China. mingchen323817@163.com.
authors: Ming
authors: M
authors: Chen
authors: Department of Clinical Laboratory, Xinqiao Hospital, Third Military Medical University, Chongqing 400037, China. guohui_chen@sina.cn.
authors: Guohui
authors: G
authors: Chen
mesh_terms: D015374:Biosensing Techniques; D004247:DNA / Q000737:chemistry / Q000302:isolation & purification*; D004261:DNA Replication / Q000235:genetics; D055664:Electrochemical Techniques; D004720:Endonucleases / Q000737:chemistry / Q000235:genetics*; D054856:G-Quadruplexes; D006427:Hemin / Q000737:chemistry / Q000235:genetics
chemical_list: D006427:Hemin; D004247:DNA; D004720:Endonucleases
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