Enhanced Detection of <i>Vibrio harveyi</i> Using a Dual-Composite DNAzyme-Based Biosensor

<i>Vibrio harveyi</i> is a serious bacterial pathogen which can infect a wide range of marine organisms, such as marine fish, invertebrates, and shrimp, in aquaculture, causing severe losses. In addition, <i>V. harveyi</i> can be transmitted through food and water, infecting...

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Main Authors: Siying Li, Shuai Zhang, Weihong Jiang, Yuying Wang, Mingwang Liu, Mingsheng Lyu, Shujun Wang
Format: Article
Language:English
Published: MDPI AG 2024-11-01
Series:Biosensors
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Online Access:https://www.mdpi.com/2079-6374/14/11/548
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author Siying Li
Shuai Zhang
Weihong Jiang
Yuying Wang
Mingwang Liu
Mingsheng Lyu
Shujun Wang
author_facet Siying Li
Shuai Zhang
Weihong Jiang
Yuying Wang
Mingwang Liu
Mingsheng Lyu
Shujun Wang
author_sort Siying Li
collection DOAJ
description <i>Vibrio harveyi</i> is a serious bacterial pathogen which can infect a wide range of marine organisms, such as marine fish, invertebrates, and shrimp, in aquaculture, causing severe losses. In addition, <i>V. harveyi</i> can be transmitted through food and water, infecting humans and posing a serious threat to public safety. Therefore, rapid and accurate detection of this pathogen is key for the prevention and control of related diseases. In this study, nine rounds of in vitro screening were conducted with Systematic Evolution of Ligands by Exponential Enrichment (SELEX) technology using unmodified DNA libraries, targeting the crude extracellular matrix (CEM) of <i>V. harveyi</i>. Two DNAzymes, named DVh<sub>1</sub> and DVh<sub>3</sub>, with high activity and specificity were obtained. Furthermore, a fluorescent biosensor with dual DNAzymes was constructed which exhibited improved detection efficiency. The sensor showed a good fluorescence response to multiple aquatic products (i.e., fish, shrimp, and shellfish) infected with <i>V. harveyi</i>, with a detection limit below 11 CFU/mL. The fluorescence signal was observed within 30 min of reaction after target addition. This simple, inexpensive, highly effective, and easy to operate DNAzymes biosensor can be used for field detection of <i>V. harveyi</i>.
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spelling doaj-art-ad60309b0d8040158ef31cf66e1dad082025-08-20T02:08:08ZengMDPI AGBiosensors2079-63742024-11-01141154810.3390/bios14110548Enhanced Detection of <i>Vibrio harveyi</i> Using a Dual-Composite DNAzyme-Based BiosensorSiying Li0Shuai Zhang1Weihong Jiang2Yuying Wang3Mingwang Liu4Mingsheng Lyu5Shujun Wang6Jiangsu Key Laboratory of Marine Bioresources and Environment/Jiangsu Key Laboratory of Marine Biotechnology, Jiangsu Ocean University, Lianyungang 222005, ChinaJiangsu Key Laboratory of Marine Bioresources and Environment/Jiangsu Key Laboratory of Marine Biotechnology, Jiangsu Ocean University, Lianyungang 222005, ChinaJiangsu Key Laboratory of Marine Bioresources and Environment/Jiangsu Key Laboratory of Marine Biotechnology, Jiangsu Ocean University, Lianyungang 222005, ChinaJiangsu Key Laboratory of Marine Bioresources and Environment/Jiangsu Key Laboratory of Marine Biotechnology, Jiangsu Ocean University, Lianyungang 222005, ChinaJiangsu Key Laboratory of Marine Bioresources and Environment/Jiangsu Key Laboratory of Marine Biotechnology, Jiangsu Ocean University, Lianyungang 222005, ChinaJiangsu Key Laboratory of Marine Bioresources and Environment/Jiangsu Key Laboratory of Marine Biotechnology, Jiangsu Ocean University, Lianyungang 222005, ChinaJiangsu Key Laboratory of Marine Bioresources and Environment/Jiangsu Key Laboratory of Marine Biotechnology, Jiangsu Ocean University, Lianyungang 222005, China<i>Vibrio harveyi</i> is a serious bacterial pathogen which can infect a wide range of marine organisms, such as marine fish, invertebrates, and shrimp, in aquaculture, causing severe losses. In addition, <i>V. harveyi</i> can be transmitted through food and water, infecting humans and posing a serious threat to public safety. Therefore, rapid and accurate detection of this pathogen is key for the prevention and control of related diseases. In this study, nine rounds of in vitro screening were conducted with Systematic Evolution of Ligands by Exponential Enrichment (SELEX) technology using unmodified DNA libraries, targeting the crude extracellular matrix (CEM) of <i>V. harveyi</i>. Two DNAzymes, named DVh<sub>1</sub> and DVh<sub>3</sub>, with high activity and specificity were obtained. Furthermore, a fluorescent biosensor with dual DNAzymes was constructed which exhibited improved detection efficiency. The sensor showed a good fluorescence response to multiple aquatic products (i.e., fish, shrimp, and shellfish) infected with <i>V. harveyi</i>, with a detection limit below 11 CFU/mL. The fluorescence signal was observed within 30 min of reaction after target addition. This simple, inexpensive, highly effective, and easy to operate DNAzymes biosensor can be used for field detection of <i>V. harveyi</i>.https://www.mdpi.com/2079-6374/14/11/548<i>Vibrio harveyi</i>dual DNAzymeSELEXfield detection
spellingShingle Siying Li
Shuai Zhang
Weihong Jiang
Yuying Wang
Mingwang Liu
Mingsheng Lyu
Shujun Wang
Enhanced Detection of <i>Vibrio harveyi</i> Using a Dual-Composite DNAzyme-Based Biosensor
Biosensors
<i>Vibrio harveyi</i>
dual DNAzyme
SELEX
field detection
title Enhanced Detection of <i>Vibrio harveyi</i> Using a Dual-Composite DNAzyme-Based Biosensor
title_full Enhanced Detection of <i>Vibrio harveyi</i> Using a Dual-Composite DNAzyme-Based Biosensor
title_fullStr Enhanced Detection of <i>Vibrio harveyi</i> Using a Dual-Composite DNAzyme-Based Biosensor
title_full_unstemmed Enhanced Detection of <i>Vibrio harveyi</i> Using a Dual-Composite DNAzyme-Based Biosensor
title_short Enhanced Detection of <i>Vibrio harveyi</i> Using a Dual-Composite DNAzyme-Based Biosensor
title_sort enhanced detection of i vibrio harveyi i using a dual composite dnazyme based biosensor
topic <i>Vibrio harveyi</i>
dual DNAzyme
SELEX
field detection
url https://www.mdpi.com/2079-6374/14/11/548
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