title: Graphene sheets, polyaniline and AuNPs based DNA sensor for electrochemical determination of BCR/ABL fusion gene with functional hairpin probe.
abstract: A sensitive and selective electrochemical DNA sensor was developed for the detection of BCR/ABL fusion gene in chronic myelogenous leukemia (CML). Firstly, graphene sheets (GS) suspension was prepared with the aid of chitosan (CS) solution and then fabricated onto the glassy carbon electrode (GCE), followed by the electro-polymerization of aniline to form the PANI layer, then, Au nanoparticles (AuNPs) were electro-deposited onto the modified GCE to immobilize the capture probes. The capture probe employed a hairpin structure and dually labeled with a 5'-SH and a 3'-biotin. After hybridization with the target DNA, hairpin structure was compelled to open and 3'-biotin was forced to stay away from the electrode surface. As a result, streptavidin-alkaline phosphatase (SA-AP) was covalently binded to the capture probe via biotin-avidin system. Reduction currents were then generated after catalyzing the hydrolysis of the electroinactive 1-naphthyl phosphate (1-NP) to 1-naphthol and monitored by differential pulse voltammetry (DPV). Under optimum conditions, the amperometric signals increased linearly with the target DNA concentrations (10 pM to 1000 pM), and the DNA sensor exhibited a detection limit as low as 2.11 pM (S/N=3) with an excellent differentiation ability, and the proposed method showed acceptable stability and reproducibility. It has been applied for assay of BCR/ABL fusion gene from real samples with satisfactory results.
authors: Key Laboratory of Laboratory Medical Diagnostics of Education, Department of Laboratory Medicine, Chongqing Medical University, No. 1 Yi Xue Yuan Road, Chongqing 400016, PR China.
authors: Li
authors: L
authors: Wang
authors: Erhui
authors: E
authors: Hua
authors: Mo
authors: M
authors: Liang
authors: Cuixia
authors: C
authors: Ma
authors: Zhangling
authors: Z
authors: Liu
authors: Shangchun
authors: S
authors: Sheng
authors: Min
authors: M
authors: Liu
authors: Guoming
authors: G
authors: Xie
authors: Wenli
authors: W
authors: Feng
mesh_terms: D000814:Aniline Compounds / Q000737:chemistry*; D015374:Biosensing Techniques / Q000379:methods; D002460:Cell Line; D004247:DNA / Q000032:analysis / Q000235:genetics*; D055664:Electrochemical Techniques / Q000379:methods*; D016044:Fusion Proteins, bcr-abl / Q000235:genetics*; D006046:Gold / Q000737:chemistry*; D006108:Graphite / Q000737:chemistry*; D006801:Humans; D053758:Nanoparticles / Q000737:chemistry* / Q000648:ultrastructure; D009693:Nucleic Acid Hybridization / Q000379:methods; D015203:Reproducibility of Results
chemical_list: D000814:Aniline Compounds; C416807:polyaniline; D006046:Gold; D006108:Graphite; D004247:DNA; D016044:Fusion Proteins, bcr-abl
keywords: AuNPs; BCR/ABL fusion gene; Electrochemical DNA sensor; Graphene sheets; Polyaniline