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A Cascade Signal Amplification Strategy for the Ultrasensitive Fluorescence Detection of Cu(2+) via λ-Exonuclease-Assisted Target Recycling with Mismatched Catalytic Hairpin Assembly
Herein, an ultrasensitive DNAzyme-based fluorescence biosensor for detecting Cu(2+) was designed using the cascade signal amplification strategy, coupling λ-exonuclease-assisted target recycling and mismatched catalytic hairpin assembly (MCHA). In the designed detection system, the target, Cu(2+), c...
Autores principales: | , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
MDPI
2023
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10605925/ https://www.ncbi.nlm.nih.gov/pubmed/37887111 http://dx.doi.org/10.3390/bios13100918 |
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author | Liu, Zhen Liu, Chen He, Liqiong Liu, Jinquan Li, Le Yang, Shengyuan Tan, Yan Liu, Xing Xiao, Xilin |
author_facet | Liu, Zhen Liu, Chen He, Liqiong Liu, Jinquan Li, Le Yang, Shengyuan Tan, Yan Liu, Xing Xiao, Xilin |
author_sort | Liu, Zhen |
collection | PubMed |
description | Herein, an ultrasensitive DNAzyme-based fluorescence biosensor for detecting Cu(2+) was designed using the cascade signal amplification strategy, coupling λ-exonuclease-assisted target recycling and mismatched catalytic hairpin assembly (MCHA). In the designed detection system, the target, Cu(2+), can activate the Cu(2+)-dependent DNAzyme to cause a cleavage reaction, releasing ssDNA (tDNA). Then, tDNA binds to hairpin DNA (H0) with an overhanging 5′-phosphorylated terminus to form dsDNA with a blunt 5′-phosphorylated terminus, which activates the dsDNA to be digested by λ-Exo and releases tDNA along with another ssDNA (iDNA). Subsequently, the iDNA initiates MCHA, which can restore the fluorescence of carboxyfluorescein (FAM) previously quenched by tetramethylrhodamine (TAMRA), resulting in a strong fluorescent signal. Furthermore, MCHA efficiently improves the signal-to-noise ratio of the detection system. More importantly, tDNA recycling can be achieved with the λ-Exo digestion reaction to release more iDNA, efficiently amplifying the fluorescent signal and further improving the sensitivity to Cu(2+) with a detection limit of 60 fM. The practical application of the developed biosensor was also demonstrated by detecting Cu(2+) in real samples, proving it to be an excellent analytical strategy for the ultrasensitive quantification of heavy metal ions in environmental water sources. |
format | Online Article Text |
id | pubmed-10605925 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2023 |
publisher | MDPI |
record_format | MEDLINE/PubMed |
spelling | pubmed-106059252023-10-28 A Cascade Signal Amplification Strategy for the Ultrasensitive Fluorescence Detection of Cu(2+) via λ-Exonuclease-Assisted Target Recycling with Mismatched Catalytic Hairpin Assembly Liu, Zhen Liu, Chen He, Liqiong Liu, Jinquan Li, Le Yang, Shengyuan Tan, Yan Liu, Xing Xiao, Xilin Biosensors (Basel) Communication Herein, an ultrasensitive DNAzyme-based fluorescence biosensor for detecting Cu(2+) was designed using the cascade signal amplification strategy, coupling λ-exonuclease-assisted target recycling and mismatched catalytic hairpin assembly (MCHA). In the designed detection system, the target, Cu(2+), can activate the Cu(2+)-dependent DNAzyme to cause a cleavage reaction, releasing ssDNA (tDNA). Then, tDNA binds to hairpin DNA (H0) with an overhanging 5′-phosphorylated terminus to form dsDNA with a blunt 5′-phosphorylated terminus, which activates the dsDNA to be digested by λ-Exo and releases tDNA along with another ssDNA (iDNA). Subsequently, the iDNA initiates MCHA, which can restore the fluorescence of carboxyfluorescein (FAM) previously quenched by tetramethylrhodamine (TAMRA), resulting in a strong fluorescent signal. Furthermore, MCHA efficiently improves the signal-to-noise ratio of the detection system. More importantly, tDNA recycling can be achieved with the λ-Exo digestion reaction to release more iDNA, efficiently amplifying the fluorescent signal and further improving the sensitivity to Cu(2+) with a detection limit of 60 fM. The practical application of the developed biosensor was also demonstrated by detecting Cu(2+) in real samples, proving it to be an excellent analytical strategy for the ultrasensitive quantification of heavy metal ions in environmental water sources. MDPI 2023-10-08 /pmc/articles/PMC10605925/ /pubmed/37887111 http://dx.doi.org/10.3390/bios13100918 Text en © 2023 by the authors. https://creativecommons.org/licenses/by/4.0/Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/). |
spellingShingle | Communication Liu, Zhen Liu, Chen He, Liqiong Liu, Jinquan Li, Le Yang, Shengyuan Tan, Yan Liu, Xing Xiao, Xilin A Cascade Signal Amplification Strategy for the Ultrasensitive Fluorescence Detection of Cu(2+) via λ-Exonuclease-Assisted Target Recycling with Mismatched Catalytic Hairpin Assembly |
title | A Cascade Signal Amplification Strategy for the Ultrasensitive Fluorescence Detection of Cu(2+) via λ-Exonuclease-Assisted Target Recycling with Mismatched Catalytic Hairpin Assembly |
title_full | A Cascade Signal Amplification Strategy for the Ultrasensitive Fluorescence Detection of Cu(2+) via λ-Exonuclease-Assisted Target Recycling with Mismatched Catalytic Hairpin Assembly |
title_fullStr | A Cascade Signal Amplification Strategy for the Ultrasensitive Fluorescence Detection of Cu(2+) via λ-Exonuclease-Assisted Target Recycling with Mismatched Catalytic Hairpin Assembly |
title_full_unstemmed | A Cascade Signal Amplification Strategy for the Ultrasensitive Fluorescence Detection of Cu(2+) via λ-Exonuclease-Assisted Target Recycling with Mismatched Catalytic Hairpin Assembly |
title_short | A Cascade Signal Amplification Strategy for the Ultrasensitive Fluorescence Detection of Cu(2+) via λ-Exonuclease-Assisted Target Recycling with Mismatched Catalytic Hairpin Assembly |
title_sort | cascade signal amplification strategy for the ultrasensitive fluorescence detection of cu(2+) via λ-exonuclease-assisted target recycling with mismatched catalytic hairpin assembly |
topic | Communication |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10605925/ https://www.ncbi.nlm.nih.gov/pubmed/37887111 http://dx.doi.org/10.3390/bios13100918 |
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