title: Dual-probe electrochemical DNA biosensor based on the "Y" junction structure and restriction endonuclease assisted cyclic enzymatic amplification for detection of double-strand DNA of PML/RARα related fusion gene.
abstract: Taking advantage of "Y" junction structure and restriction endonuclease assisted cyclic enzymatic amplification, a dual-probe electrochemical DNA (DE-DNA) biosensor was designed to detect double-stranded DNA (dsDNA) of acute promyelocytic leukemia (APL) related gene. Two groups of detection probes were designed, and each group was composed of a biotinylated capture probe and an assisted probe. They were separately complementary with two strands of target dsDNA in order to prevent the reannealing of the two separate strands from target dsDNA. First, thiol functionalized capture probes (C1 and C2) were severally assembled onto two different gold electrodes, followed by hybridizing with target dsDNA (S1a-S1b) and assistant probes to form two Y-junction-structure ternary complexes. Subsequently, restriction sites on the ternary complexes were digested by Rsa I, which can release S1a, S1b and biotins from the electrode surfaces. Meanwhile, the released S1a and S1b can further hybridize with the unhybridized corresponding detection probes and then initiate another new hybridization-cleavage-separation cycle. Finally, the current signals were produced by the enzyme-catalyzed reaction of streptavidin-horse reddish peroxidase (streptavidin-HRP). The distinct difference in current signals between different sequences allowed detection of target dsDNA down to a low detection limit of 47 fM and presented excellent specificity with discriminating only a single-base mismatched dsDNA sequence. Moreover, this biosensor was also used for assay of polymerase chain reaction (PCR) samples with satisfactory results. According to the results, the power of the DE-DNA biosensor as a promising tool for the detection of APL and other diseases.
authors.affiliation: Department of Pharmaceutical Analysis, Faculty of Pharmacy, Fujian Medical University, Fuzhou 350004, China; Department of Pharmacy, the First Affiliated Hospital of Xiamen University, Xiamen 361003, China.
authors.forename: Kun
authors.initials: K
authors.lastname: Wang
authors.affiliation: Department of Pharmaceutical Analysis, Faculty of Pharmacy, Fujian Medical University, Fuzhou 350004, China; Nano Biomedical Technology Research Center, Fujian Medical University, Fuzhou 350004, China.
authors.forename: Yun
authors.initials: Y
authors.lastname: Lei
authors.affiliation: Department of Pharmaceutical Analysis, Faculty of Pharmacy, Fujian Medical University, Fuzhou 350004, China; Department of Orthopaedics, the First Affiliated Hospital of Fujian Medical University, Fuzhou 350004 China; Nano Biomedical Technology Research Center, Fujian Medical University, Fuzhou 350004, China.
authors.forename: Guang-Xian
authors.initials: GX
authors.lastname: Zhong
authors.affiliation: Department of Pharmaceutical Analysis, Faculty of Pharmacy, Fujian Medical University, Fuzhou 350004, China; Nano Biomedical Technology Research Center, Fujian Medical University, Fuzhou 350004, China.
authors.forename: Yan-Jie
authors.initials: YJ
authors.lastname: Zheng
authors.affiliation: Department of Pharmacy, the First Affiliated Hospital of Xiamen University, Xiamen 361003, China.
authors.forename: Zhou-Liang
authors.initials: ZL
authors.lastname: Sun
authors.affiliation: Department of Pharmaceutical Analysis, Faculty of Pharmacy, Fujian Medical University, Fuzhou 350004, China; Nano Biomedical Technology Research Center, Fujian Medical University, Fuzhou 350004, China.
authors.forename: Hua-Ping
authors.initials: HP
authors.lastname: Peng
authors.affiliation: Department of Pharmaceutical Analysis, Faculty of Pharmacy, Fujian Medical University, Fuzhou 350004, China; Nano Biomedical Technology Research Center, Fujian Medical University, Fuzhou 350004, China.
authors.forename: Wei
authors.initials: W
authors.lastname: Chen
authors.affiliation: Department of Pharmaceutical Analysis, Faculty of Pharmacy, Fujian Medical University, Fuzhou 350004, China; Nano Biomedical Technology Research Center, Fujian Medical University, Fuzhou 350004, China. Electronic address: ailinliu@mail.fjmu.edu.cn.
authors.forename: Ai-Lin
authors.initials: AL
authors.lastname: Liu
authors.affiliation: Fujian Institute of Hematology, the Affiliated Union Hospital of Fujian Medical University, Fuzhou 350000, China; Nano Biomedical Technology Research Center, Fujian Medical University, Fuzhou 350004, China. Electronic address: chenyz@mail.fjmu.edu.cn.
authors.forename: Yuan-Zhong
authors.initials: YZ
authors.lastname: Chen
authors.affiliation: Department of Pharmaceutical Analysis, Faculty of Pharmacy, Fujian Medical University, Fuzhou 350004, China; Nano Biomedical Technology Research Center, Fujian Medical University, Fuzhou 350004, China. Electronic address: xhl1963@sina.com.
authors.forename: Xin-Hua
authors.initials: XH
authors.lastname: Lin
mesh_terms: D001483:Base Sequence; D003217:Conductometry / Q000295:instrumentation*; D004247:DNA / Q000032:analysis / Q000737:chemistry / Q000235:genetics*; D015342:DNA Probes / Q000737:chemistry / Q000235:genetics*; D004262:DNA Restriction Enzymes / Q000737:chemistry / Q000235:genetics; D004867:Equipment Design; D019544:Equipment Failure Analysis; D008969:Molecular Sequence Data; D009687:Nuclear Proteins / Q000032:analysis / Q000737:chemistry / Q000235:genetics*; D021141:Nucleic Acid Amplification Techniques / Q000295:instrumentation; D020411:Oligonucleotide Array Sequence Analysis / Q000295:instrumentation*; D000071156:Promyelocytic Leukemia Protein; D018168:Receptors, Retinoic Acid / Q000032:analysis / Q000737:chemistry / Q000235:genetics*; D011993:Recombinant Fusion Proteins / Q000032:analysis / Q000235:genetics; D015203:Reproducibility of Results; D000072482:Retinoic Acid Receptor alpha; D012680:Sensitivity and Specificity; D017422:Sequence Analysis, DNA / Q000295:instrumentation; D014157:Transcription Factors / Q000032:analysis / Q000737:chemistry / Q000235:genetics*; D025521:Tumor Suppressor Proteins / Q000032:analysis / Q000737:chemistry / Q000235:genetics*
chemical_list: D015342:DNA Probes; D009687:Nuclear Proteins; D000071156:Promyelocytic Leukemia Protein; C000607071:RARA protein, human; D018168:Receptors, Retinoic Acid; D011993:Recombinant Fusion Proteins; D000072482:Retinoic Acid Receptor alpha; D014157:Transcription Factors; D025521:Tumor Suppressor Proteins; C070576:PML protein, human; D004247:DNA; D004262:DNA Restriction Enzymes
keywords: Dual-probe electrochemical DNA biosensor; PML/RARα; Restriction endonuclease; “Y” junction structure