title: An enhanced chemiluminescence resonance energy transfer system based on target recycling G-guadruplexes/hemin DNAzyme catalysis and its application in ultrasensitive detection of DNA.
abstract: An enhanced chemiluminescence resonance energy transfer (CRET) system based on target recycling G-guadruplexes/hemin DNAzyme catalysis was developed for ultrasensitive detection of DNA. CRET system consists of luminol as chemiluminescent donor, and fluorescein isothiocyanate (FITC) as acceptor. The sensitive detection was achieved by using the system consisted of G-riched DNA, blocker DNA, and the Nb.BbvCI biocatalyst. Upon addition of target DNA to the system, target DNA hybridizes with the quasi-circular DNA structure, and forms a DNA duplex. The formation of DNA duplex triggers selective enzymatic cleavage of quasi-circular DNA by Nb.BbvCI, resulting in the release of target DNA and two G-riched DNAzyme segments. Released target DNA then hybridizes with another quasi-circular DNA structure to initiate the cleavage of the quasi-circular DNA structure. Eventually, each target DNA can go through many cycles, resulting in the digestion of many quasi-circular DNA structures, generating many G-riched DNAzyme segments. G-riched DNAzyme segment products assemble with hemin to form stable hemin/G-quadruplexes that exhibit peroxidase-like activity which can catalyze the oxidation of luminol by H2O2 to produce CL signals. In the presence of FITC, CL of luminol can excite FITC molecules, and thus produced CRET between the luminol and FITC. This unique analysis strategy gives a detection limit down to 80 fM, which is at least four orders of magnitude lower than that of unamplified DNA detection methods.
authors: Key Laboratory for the Chemistry and Molecular Engineering of Medicinal Resources (Ministry of Education), College of Chemistry and Pharmacy, Guangxi Normal University, Guilin 541004, China.
authors: Jia
authors: J
authors: Chen
authors: Key Laboratory for the Chemistry and Molecular Engineering of Medicinal Resources (Ministry of Education), College of Chemistry and Pharmacy, Guangxi Normal University, Guilin 541004, China.
authors: Yong
authors: Y
authors: Huang
authors: Department of Chemistry, Lomonosov Moscow State University, Moscow 119991, Russia.
authors: Marina
authors: M
authors: Vdovenko
authors: Department of Chemistry, Lomonosov Moscow State University, Moscow 119991, Russia.
authors: Ivan Yu
authors: IY
authors: Sakharov
authors: Key Laboratory for the Chemistry and Molecular Engineering of Medicinal Resources (Ministry of Education), College of Chemistry and Pharmacy, Guangxi Normal University, Guilin 541004, China.
authors: Guifa
authors: G
authors: Su
authors: Key Laboratory for the Chemistry and Molecular Engineering of Medicinal Resources (Ministry of Education), College of Chemistry and Pharmacy, Guangxi Normal University, Guilin 541004, China. Electronic address: zhaoshulin001@163.com.
authors: Shulin
authors: S
authors: Zhao
mesh_terms: D015374:Biosensing Techniques / Q000379:methods*; D002384:Catalysis; D004247:DNA / Q000032:analysis*; D021881:DNA, Catalytic / Q000737:chemistry*; D003911:Dextrans; D004735:Energy Transfer; D016650:Fluorescein-5-isothiocyanate / Q000031:analogs & derivatives; D054856:G-Quadruplexes; D006427:Hemin / Q000737:chemistry*; D006801:Humans; D006861:Hydrogen Peroxide / Q000737:chemistry*; D049449:Luminescence; D008163:Luminescent Measurements / Q000379:methods*; D008165:Luminol / Q000737:chemistry; D009693:Nucleic Acid Hybridization
chemical_list: D021881:DNA, Catalytic; D003911:Dextrans; C015219:fluorescein isothiocyanate dextran; D008165:Luminol; D006427:Hemin; D004247:DNA; D006861:Hydrogen Peroxide; D016650:Fluorescein-5-isothiocyanate
keywords: Chemiluminescence resonance energy transfer; DNA detection; G-quadruplexes/hemin DNAzyme catalysis; Target recycling