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A cascade signal amplification strategy for surface enhanced Raman spectroscopy detection of thrombin based on DNAzyme assistant DNA recycling and rolling circle amplification.
Biosens Bioelectron. 2015 Apr 15; 66:423-30.BB

Abstract

A sensitive protocol for surface enhanced Raman spectroscopy (SERS) detection of thrombin is designed with R6G-Ag NPs as a signal tag by combining DNAzyme assistant DNA recycling and rolling circle amplification (RCA). Molecular beacon (MB) as recognition probe immobilizes on the glass slides and performs the amplification procedure. After thrombin-induced structure-switching DNA hairpins of probe 1, the DNAzyme is liberated from the caged structure, which hybridizes with the MB for cleavage of the MB in the presence of cofactor Zn(2+) and initiates the DNA recycling process, leading to the cleavage of a large number of MB and the generation of numerous primers for triggering RCA reaction. The long amplified RCA product which contained hundreds of tandem-repeat sequences, which can bind with oligonucleotide functionalized Ag NPs reporters. The attached signal tags can be easily read out by SERS. Because of the cascade signal amplification, these newly designed protocols provides a sensitive SERS detection of thrombin down to the femolar level (2.3fM) with a linear range of 5 orders of magnitude (from 10(-14) to 10(-9)M) and have high selectivity toward its target protein. The proposed method is expected to be a good clinical tool for the diagnosis of a thrombotic disease.

Authors+Show Affiliations

Jiangsu Key Laboratory of New Drug Research and Clinical Pharmacy, Xuzhou Medical College, 221004 Xuzhou, China. Electronic address: jsxzgfl@sina.com.Jiangsu Key Laboratory of New Drug Research and Clinical Pharmacy, Xuzhou Medical College, 221004 Xuzhou, China.Jiangsu Key Laboratory of New Drug Research and Clinical Pharmacy, Xuzhou Medical College, 221004 Xuzhou, China.Jiangsu Key Laboratory of New Drug Research and Clinical Pharmacy, Xuzhou Medical College, 221004 Xuzhou, China.Jiangsu Key Laboratory of New Drug Research and Clinical Pharmacy, Xuzhou Medical College, 221004 Xuzhou, China.Jiangsu Key Laboratory of New Drug Research and Clinical Pharmacy, Xuzhou Medical College, 221004 Xuzhou, China.

Pub Type(s)

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

Language

eng

PubMed ID

25497982

Citation

Gao, Fenglei, et al. "A Cascade Signal Amplification Strategy for Surface Enhanced Raman Spectroscopy Detection of Thrombin Based On DNAzyme Assistant DNA Recycling and Rolling Circle Amplification." Biosensors & Bioelectronics, vol. 66, 2015, pp. 423-30.
Gao F, Du L, Tang D, et al. A cascade signal amplification strategy for surface enhanced Raman spectroscopy detection of thrombin based on DNAzyme assistant DNA recycling and rolling circle amplification. Biosens Bioelectron. 2015;66:423-30.
Gao, F., Du, L., Tang, D., Lu, Y., Zhang, Y., & Zhang, L. (2015). A cascade signal amplification strategy for surface enhanced Raman spectroscopy detection of thrombin based on DNAzyme assistant DNA recycling and rolling circle amplification. Biosensors & Bioelectronics, 66, 423-30. https://doi.org/10.1016/j.bios.2014.12.006
Gao F, et al. A Cascade Signal Amplification Strategy for Surface Enhanced Raman Spectroscopy Detection of Thrombin Based On DNAzyme Assistant DNA Recycling and Rolling Circle Amplification. Biosens Bioelectron. 2015 Apr 15;66:423-30. PubMed PMID: 25497982.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - A cascade signal amplification strategy for surface enhanced Raman spectroscopy detection of thrombin based on DNAzyme assistant DNA recycling and rolling circle amplification. AU - Gao,Fenglei, AU - Du,Lili, AU - Tang,Daoquan, AU - Lu,Yao, AU - Zhang,Yanzhuo, AU - Zhang,Lixian, Y1 - 2014/12/03/ PY - 2014/09/01/received PY - 2014/11/11/revised PY - 2014/12/01/accepted PY - 2014/12/16/entrez PY - 2014/12/17/pubmed PY - 2015/9/1/medline KW - DNAzyme KW - Rolling circle amplification KW - Signal amplification KW - Surface enhanced Raman spectroscopy SP - 423 EP - 30 JF - Biosensors & bioelectronics JO - Biosens Bioelectron VL - 66 N2 - A sensitive protocol for surface enhanced Raman spectroscopy (SERS) detection of thrombin is designed with R6G-Ag NPs as a signal tag by combining DNAzyme assistant DNA recycling and rolling circle amplification (RCA). Molecular beacon (MB) as recognition probe immobilizes on the glass slides and performs the amplification procedure. After thrombin-induced structure-switching DNA hairpins of probe 1, the DNAzyme is liberated from the caged structure, which hybridizes with the MB for cleavage of the MB in the presence of cofactor Zn(2+) and initiates the DNA recycling process, leading to the cleavage of a large number of MB and the generation of numerous primers for triggering RCA reaction. The long amplified RCA product which contained hundreds of tandem-repeat sequences, which can bind with oligonucleotide functionalized Ag NPs reporters. The attached signal tags can be easily read out by SERS. Because of the cascade signal amplification, these newly designed protocols provides a sensitive SERS detection of thrombin down to the femolar level (2.3fM) with a linear range of 5 orders of magnitude (from 10(-14) to 10(-9)M) and have high selectivity toward its target protein. The proposed method is expected to be a good clinical tool for the diagnosis of a thrombotic disease. SN - 1873-4235 UR - https://www.unboundmedicine.com/medline/citation/25497982/A_cascade_signal_amplification_strategy_for_surface_enhanced_Raman_spectroscopy_detection_of_thrombin_based_on_DNAzyme_assistant_DNA_recycling_and_rolling_circle_amplification_ DB - PRIME DP - Unbound Medicine ER -