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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MDPI AG
2024-11-01
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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 |
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| 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>. |
| format | Article |
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| institution | OA Journals |
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| language | English |
| publishDate | 2024-11-01 |
| publisher | MDPI AG |
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| series | Biosensors |
| 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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