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Hyperbranched rolling circle amplification based electrochemiluminescence aptasensor for ultrasensitive detection of thrombin.
Biosens Bioelectron. 2015 Jan 15; 63:166-171.BB

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+Show Affiliations

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.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.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.College of Environment and Resources, Fuzhou University, Fuzhou, Fujian 350116, China. Electronic address: lixiaojuan@fzu.edu.cn.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.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.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.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.

Pub Type(s)

Journal Article
Research Support, Non-U.S. Gov't

Language

eng

PubMed ID

25086328

Citation

Jin, Guixiao, et al. "Hyperbranched Rolling Circle Amplification Based Electrochemiluminescence Aptasensor for Ultrasensitive Detection of Thrombin." Biosensors & Bioelectronics, vol. 63, 2015, pp. 166-171.
Jin G, Wang C, Yang L, et al. Hyperbranched rolling circle amplification based electrochemiluminescence aptasensor for ultrasensitive detection of thrombin. Biosens Bioelectron. 2015;63:166-171.
Jin, G., Wang, C., Yang, L., Li, X., Guo, L., Qiu, B., Lin, Z., & Chen, G. (2015). Hyperbranched rolling circle amplification based electrochemiluminescence aptasensor for ultrasensitive detection of thrombin. Biosensors & Bioelectronics, 63, 166-171. https://doi.org/10.1016/j.bios.2014.07.033
Jin G, et al. Hyperbranched Rolling Circle Amplification Based Electrochemiluminescence Aptasensor for Ultrasensitive Detection of Thrombin. Biosens Bioelectron. 2015 Jan 15;63:166-171. PubMed PMID: 25086328.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Hyperbranched rolling circle amplification based electrochemiluminescence aptasensor for ultrasensitive detection of thrombin. AU - Jin,Guixiao, AU - Wang,Chunmei, AU - Yang,Linlin, AU - Li,Xiaojuan, AU - Guo,Longhua, AU - Qiu,Bin, AU - Lin,Zhenyu, AU - Chen,Guonan, Y1 - 2014/07/22/ PY - 2014/05/22/received PY - 2014/07/09/revised PY - 2014/07/17/accepted PY - 2014/8/3/entrez PY - 2014/8/3/pubmed PY - 2015/9/9/medline KW - Aptamer KW - Electrochemiluminescence KW - HRCA KW - Thrombin SP - 166 EP - 171 JF - Biosensors & bioelectronics JO - Biosens Bioelectron VL - 63 N2 - 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. SN - 1873-4235 UR - https://www.unboundmedicine.com/medline/citation/25086328/Hyperbranched_rolling_circle_amplification_based_electrochemiluminescence_aptasensor_for_ultrasensitive_detection_of_thrombin_ DB - PRIME DP - Unbound Medicine ER -