Cascade Fluorescent Sensors Based on Isothermal Signal Amplification for the Detection of Mercury and Silver Ions

In this study, novel fluorescent DNA biosensors for mercury (Hg<sup>2+</sup>) and silver (Ag<sup>+</sup>) ions were developed based on thymine (T)- and cytosine (C)-rich recognition elements in combination with exonuclease III and a mismatch-catalyzed hairpin assembly (MCHA)-...

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Main Authors: Zhen Liu, Xing Liu, Jie Sun, Xilin Xiao
Format: Article
Language:English
Published: MDPI AG 2025-03-01
Series:Biosensors
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Online Access:https://www.mdpi.com/2079-6374/15/4/213
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author Zhen Liu
Xing Liu
Jie Sun
Xilin Xiao
author_facet Zhen Liu
Xing Liu
Jie Sun
Xilin Xiao
author_sort Zhen Liu
collection DOAJ
description In this study, novel fluorescent DNA biosensors for mercury (Hg<sup>2+</sup>) and silver (Ag<sup>+</sup>) ions were developed based on thymine (T)- and cytosine (C)-rich recognition elements in combination with exonuclease III and a mismatch-catalyzed hairpin assembly (MCHA)-based cascade isothermal signal-amplification strategy. In the presence of the respective target analytes, the recognition element terminals form so-called T-Hg<sup>2+</sup>-T or C-Ag<sup>+</sup>-C structures, resulting in cleavage by Exo III and the release of the trigger strand for MCHA. This binds to the H1 hairpin, which is fluorescently labeled with carboxyfluorescein (FAM) and tetramethylrhodamine (TAMRA), disrupting fluorescence resonance energy transfer between them and, thus, restoring FAM fluorescence, generating a strong signal at 520 nm. The linear range of the Hg<sup>2+</sup> sensor is 0.5 to 3 pM, with a detection limit of 0.07 pM. The recovery range in actual spiked water samples is between 98.5% and 105.2%, with a relative standard deviation (RSD) ranging from 2.0% to 4.2%. The linear range of the Ag<sup>+</sup> sensor is 10 to 90 pM, with a detection limit of 7.6 pM. The recovery range in actual spiked water samples is between 96.2% and 104.1%, with an RSD ranging from 3.2% to 6.3%. The cascade isothermal signal amplification strategy effectively enhances sensor sensitivity, while MCHA decreases the false-positive rate. The aptamer sensor exhibits high specificity, is resistant to interference, and can be used for the detection of Hg<sup>2+</sup> and Ag<sup>+</sup> in environmental water samples.
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spelling doaj-art-edf2846e5a4f45e1a921f30b516b0c492025-08-20T02:28:40ZengMDPI AGBiosensors2079-63742025-03-0115421310.3390/bios15040213Cascade Fluorescent Sensors Based on Isothermal Signal Amplification for the Detection of Mercury and Silver IonsZhen Liu0Xing Liu1Jie Sun2Xilin Xiao3School of Public Health, Hengyang School of Medicine, University of South China, Hengyang 421001, ChinaSchool of Public Health, Hengyang School of Medicine, University of South China, Hengyang 421001, ChinaSchool of Public Health, Hengyang School of Medicine, University of South China, Hengyang 421001, ChinaSchool of Public Health, Hengyang School of Medicine, University of South China, Hengyang 421001, ChinaIn this study, novel fluorescent DNA biosensors for mercury (Hg<sup>2+</sup>) and silver (Ag<sup>+</sup>) ions were developed based on thymine (T)- and cytosine (C)-rich recognition elements in combination with exonuclease III and a mismatch-catalyzed hairpin assembly (MCHA)-based cascade isothermal signal-amplification strategy. In the presence of the respective target analytes, the recognition element terminals form so-called T-Hg<sup>2+</sup>-T or C-Ag<sup>+</sup>-C structures, resulting in cleavage by Exo III and the release of the trigger strand for MCHA. This binds to the H1 hairpin, which is fluorescently labeled with carboxyfluorescein (FAM) and tetramethylrhodamine (TAMRA), disrupting fluorescence resonance energy transfer between them and, thus, restoring FAM fluorescence, generating a strong signal at 520 nm. The linear range of the Hg<sup>2+</sup> sensor is 0.5 to 3 pM, with a detection limit of 0.07 pM. The recovery range in actual spiked water samples is between 98.5% and 105.2%, with a relative standard deviation (RSD) ranging from 2.0% to 4.2%. The linear range of the Ag<sup>+</sup> sensor is 10 to 90 pM, with a detection limit of 7.6 pM. The recovery range in actual spiked water samples is between 96.2% and 104.1%, with an RSD ranging from 3.2% to 6.3%. The cascade isothermal signal amplification strategy effectively enhances sensor sensitivity, while MCHA decreases the false-positive rate. The aptamer sensor exhibits high specificity, is resistant to interference, and can be used for the detection of Hg<sup>2+</sup> and Ag<sup>+</sup> in environmental water samples.https://www.mdpi.com/2079-6374/15/4/213fluorescent sensormercury ion detectionsilver ion detectioncascade isothermal signal amplificationcatalytic hairpin assembly
spellingShingle Zhen Liu
Xing Liu
Jie Sun
Xilin Xiao
Cascade Fluorescent Sensors Based on Isothermal Signal Amplification for the Detection of Mercury and Silver Ions
Biosensors
fluorescent sensor
mercury ion detection
silver ion detection
cascade isothermal signal amplification
catalytic hairpin assembly
title Cascade Fluorescent Sensors Based on Isothermal Signal Amplification for the Detection of Mercury and Silver Ions
title_full Cascade Fluorescent Sensors Based on Isothermal Signal Amplification for the Detection of Mercury and Silver Ions
title_fullStr Cascade Fluorescent Sensors Based on Isothermal Signal Amplification for the Detection of Mercury and Silver Ions
title_full_unstemmed Cascade Fluorescent Sensors Based on Isothermal Signal Amplification for the Detection of Mercury and Silver Ions
title_short Cascade Fluorescent Sensors Based on Isothermal Signal Amplification for the Detection of Mercury and Silver Ions
title_sort cascade fluorescent sensors based on isothermal signal amplification for the detection of mercury and silver ions
topic fluorescent sensor
mercury ion detection
silver ion detection
cascade isothermal signal amplification
catalytic hairpin assembly
url https://www.mdpi.com/2079-6374/15/4/213
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AT xingliu cascadefluorescentsensorsbasedonisothermalsignalamplificationforthedetectionofmercuryandsilverions
AT jiesun cascadefluorescentsensorsbasedonisothermalsignalamplificationforthedetectionofmercuryandsilverions
AT xilinxiao cascadefluorescentsensorsbasedonisothermalsignalamplificationforthedetectionofmercuryandsilverions