title: Hyperbranched rolling circle amplification based electrochemiluminescence aptasensor for ultrasensitive detection of thrombin.
abstract: An ultrasensitive electrochemiluminescence (ECL) aptamer sensor for protein (thrombin as an example) detection based on hyperbranched rolling circle amplification (HRCA) had been developed. A complementary single-strand DNA (CDNA) of the thrombin aptamer had been modified on the gold electrode firstly, and then hybridized with thrombin aptamer to make the aptamer immobilized on the electrode surface, in the presence of thrombin, aptamer-thrombin bioaffinity complexes formed and made thrombin aptamer leave the electrode surface. Thus, the linear padlock probe hybridized with the free CDNA on the electrode surface and circularized by Escherichia coli DNA ligase. Subsequently, the linear padlock probe was served as a template for the initiation of HRCA reaction, and a lot of dsDNA modified on the electrode surface. Then Ru(phen)₃²⁺ (acted as the ECL indicator) intercalates specifically into double-stranded DNA (dsDNA) grooves to generate ECL signal. The ECL intensity of the system has a linear relationship with thrombin concentration in the range of 3.0-300 aM with a detection limit of 1.2 aM (S/N=3). The proposed method combines the high sensitivity of ECL, exponential ampliﬁcation of HRCA for signal enhancement and high selectivity of aptamer.
authors.affiliation: MOE Key Laboratory of Analysis and Detection for Food Safety, Fujian Provincial Key Laboratory of Analysis and Detection Technology for Food Safety, Institute of Nanomedicine and Nanobiosensing, College of Chemistry, Fuzhou University, Fuzhou, Fujian 350116, China.
authors.forename: Guixiao
authors.initials: G
authors.lastname: Jin
authors.affiliation: MOE Key Laboratory of Analysis and Detection for Food Safety, Fujian Provincial Key Laboratory of Analysis and Detection Technology for Food Safety, Institute of Nanomedicine and Nanobiosensing, College of Chemistry, Fuzhou University, Fuzhou, Fujian 350116, China.
authors.forename: Chunmei
authors.initials: C
authors.lastname: Wang
authors.affiliation: MOE Key Laboratory of Analysis and Detection for Food Safety, Fujian Provincial Key Laboratory of Analysis and Detection Technology for Food Safety, Institute of Nanomedicine and Nanobiosensing, College of Chemistry, Fuzhou University, Fuzhou, Fujian 350116, China.
authors.forename: Linlin
authors.initials: L
authors.lastname: Yang
authors.affiliation: College of Environment and Resources, Fuzhou University, Fuzhou, Fujian 350116, China. Electronic address: lixiaojuan@fzu.edu.cn.
authors.forename: Xiaojuan
authors.initials: X
authors.lastname: Li
authors.affiliation: MOE Key Laboratory of Analysis and Detection for Food Safety, Fujian Provincial Key Laboratory of Analysis and Detection Technology for Food Safety, Institute of Nanomedicine and Nanobiosensing, College of Chemistry, Fuzhou University, Fuzhou, Fujian 350116, China.
authors.forename: Longhua
authors.initials: L
authors.lastname: Guo
authors.affiliation: MOE Key Laboratory of Analysis and Detection for Food Safety, Fujian Provincial Key Laboratory of Analysis and Detection Technology for Food Safety, Institute of Nanomedicine and Nanobiosensing, College of Chemistry, Fuzhou University, Fuzhou, Fujian 350116, China.
authors.forename: Bin
authors.initials: B
authors.lastname: Qiu
authors.affiliation: MOE Key Laboratory of Analysis and Detection for Food Safety, Fujian Provincial Key Laboratory of Analysis and Detection Technology for Food Safety, Institute of Nanomedicine and Nanobiosensing, College of Chemistry, Fuzhou University, Fuzhou, Fujian 350116, China. Electronic address: zylin@fzu.edu.cn.
authors.forename: Zhenyu
authors.initials: Z
authors.lastname: Lin
authors.affiliation: MOE Key Laboratory of Analysis and Detection for Food Safety, Fujian Provincial Key Laboratory of Analysis and Detection Technology for Food Safety, Institute of Nanomedicine and Nanobiosensing, College of Chemistry, Fuzhou University, Fuzhou, Fujian 350116, China.
authors.forename: Guonan
authors.initials: G
authors.lastname: Chen
mesh_terms: D052157:Aptamers, Nucleotide / Q000737:chemistry / Q000235:genetics*; D015374:Biosensing Techniques / Q000295:instrumentation*; D001774:Blood Chemical Analysis / Q000295:instrumentation*; D003217:Conductometry / Q000295:instrumentation; D004867:Equipment Design; D019544:Equipment Failure Analysis; D006801:Humans; D008163:Luminescent Measurements / Q000295:instrumentation*; D021141:Nucleic Acid Amplification Techniques / Q000295:instrumentation*; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D013917:Thrombin / Q000032:analysis* / Q000737:chemistry / Q000235:genetics
chemical_list: D052157:Aptamers, Nucleotide; D013917:Thrombin
keywords: Aptamer; Electrochemiluminescence; HRCA; Thrombin