Na Ying, Yuan Wang, Bo Qin, Yanqing Wu, Zitong Wang, Huijuan Chen, Xuefeng Song, Zhixing Su, Wenhong Fang
{"title":"基于重组酶聚合酶扩增和链置换反应的肝芽胞核孢子虫侧流核酸检测。","authors":"Na Ying, Yuan Wang, Bo Qin, Yanqing Wu, Zitong Wang, Huijuan Chen, Xuefeng Song, Zhixing Su, Wenhong Fang","doi":"10.3354/dao03847","DOIUrl":null,"url":null,"abstract":"<p><p>The incidence of Ecytonucleospora hepatopenaei (EHP) infections in farmed shrimp has increased markedly in recent years, resulting in significant economic losses for the global shrimp farming industry. The lack of an efficacious drug for EHP infection has led to the development of a strategy based on the timely screening and elimination of EHP-carrying shrimp seeds as a means of preventing financial loss. This strategy requires portable, accurate and rapid detection methods for EHP, especially when applied to sites such as farms. However, the current lack of user-friendly devices capable of real-time detection under field conditions represents a significant challenge in the implementation of this strategy. In this study, an isothermal amplification nucleic acid biosensor for EHP detection was developed. The biosensor targeted the spore wall protein gene of EHP and amplified the target gene by recombinase polymerase amplification (RPA) combined with strand displacement reaction (SDR). The amplified products were applied on gold nanoparticle-based lateral flow nucleic acid strips (LFNAS) for visual signal conversion. The limit of detection of the SDR-RPA-LFNAS assay was 7 copies reaction-1, and the entire process could be completed in 30 min without cross-reaction. In contrast to existing conventional RPA-related detection methods, the introduction of SDR, which is used to eliminate the background signal produced by long primers, avoids the use of endonucleases and reduces costs. Moreover, the biosensor is straightforward to operate and does not require the use of expensive machinery, rendering it more suitable for the in situ detection of EHP in shrimp farms or aquaculture facilities.</p>","PeriodicalId":11252,"journal":{"name":"Diseases of aquatic organisms","volume":"162 ","pages":"17-26"},"PeriodicalIF":1.1000,"publicationDate":"2025-04-17","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"0","resultStr":"{\"title\":\"Lateral flow nucleic acid assay for Ecytonucleospora hepatopenaei based on recombinase polymerase amplification and strand displacement reaction.\",\"authors\":\"Na Ying, Yuan Wang, Bo Qin, Yanqing Wu, Zitong Wang, Huijuan Chen, Xuefeng Song, Zhixing Su, Wenhong Fang\",\"doi\":\"10.3354/dao03847\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"<p><p>The incidence of Ecytonucleospora hepatopenaei (EHP) infections in farmed shrimp has increased markedly in recent years, resulting in significant economic losses for the global shrimp farming industry. The lack of an efficacious drug for EHP infection has led to the development of a strategy based on the timely screening and elimination of EHP-carrying shrimp seeds as a means of preventing financial loss. This strategy requires portable, accurate and rapid detection methods for EHP, especially when applied to sites such as farms. However, the current lack of user-friendly devices capable of real-time detection under field conditions represents a significant challenge in the implementation of this strategy. In this study, an isothermal amplification nucleic acid biosensor for EHP detection was developed. The biosensor targeted the spore wall protein gene of EHP and amplified the target gene by recombinase polymerase amplification (RPA) combined with strand displacement reaction (SDR). The amplified products were applied on gold nanoparticle-based lateral flow nucleic acid strips (LFNAS) for visual signal conversion. The limit of detection of the SDR-RPA-LFNAS assay was 7 copies reaction-1, and the entire process could be completed in 30 min without cross-reaction. In contrast to existing conventional RPA-related detection methods, the introduction of SDR, which is used to eliminate the background signal produced by long primers, avoids the use of endonucleases and reduces costs. Moreover, the biosensor is straightforward to operate and does not require the use of expensive machinery, rendering it more suitable for the in situ detection of EHP in shrimp farms or aquaculture facilities.</p>\",\"PeriodicalId\":11252,\"journal\":{\"name\":\"Diseases of aquatic organisms\",\"volume\":\"162 \",\"pages\":\"17-26\"},\"PeriodicalIF\":1.1000,\"publicationDate\":\"2025-04-17\",\"publicationTypes\":\"Journal Article\",\"fieldsOfStudy\":null,\"isOpenAccess\":false,\"openAccessPdf\":\"\",\"citationCount\":\"0\",\"resultStr\":null,\"platform\":\"Semanticscholar\",\"paperid\":null,\"PeriodicalName\":\"Diseases of aquatic organisms\",\"FirstCategoryId\":\"97\",\"ListUrlMain\":\"https://doi.org/10.3354/dao03847\",\"RegionNum\":4,\"RegionCategory\":\"农林科学\",\"ArticlePicture\":[],\"TitleCN\":null,\"AbstractTextCN\":null,\"PMCID\":null,\"EPubDate\":\"\",\"PubModel\":\"\",\"JCR\":\"Q3\",\"JCRName\":\"FISHERIES\",\"Score\":null,\"Total\":0}","platform":"Semanticscholar","paperid":null,"PeriodicalName":"Diseases of aquatic organisms","FirstCategoryId":"97","ListUrlMain":"https://doi.org/10.3354/dao03847","RegionNum":4,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q3","JCRName":"FISHERIES","Score":null,"Total":0}
Lateral flow nucleic acid assay for Ecytonucleospora hepatopenaei based on recombinase polymerase amplification and strand displacement reaction.
The incidence of Ecytonucleospora hepatopenaei (EHP) infections in farmed shrimp has increased markedly in recent years, resulting in significant economic losses for the global shrimp farming industry. The lack of an efficacious drug for EHP infection has led to the development of a strategy based on the timely screening and elimination of EHP-carrying shrimp seeds as a means of preventing financial loss. This strategy requires portable, accurate and rapid detection methods for EHP, especially when applied to sites such as farms. However, the current lack of user-friendly devices capable of real-time detection under field conditions represents a significant challenge in the implementation of this strategy. In this study, an isothermal amplification nucleic acid biosensor for EHP detection was developed. The biosensor targeted the spore wall protein gene of EHP and amplified the target gene by recombinase polymerase amplification (RPA) combined with strand displacement reaction (SDR). The amplified products were applied on gold nanoparticle-based lateral flow nucleic acid strips (LFNAS) for visual signal conversion. The limit of detection of the SDR-RPA-LFNAS assay was 7 copies reaction-1, and the entire process could be completed in 30 min without cross-reaction. In contrast to existing conventional RPA-related detection methods, the introduction of SDR, which is used to eliminate the background signal produced by long primers, avoids the use of endonucleases and reduces costs. Moreover, the biosensor is straightforward to operate and does not require the use of expensive machinery, rendering it more suitable for the in situ detection of EHP in shrimp farms or aquaculture facilities.
期刊介绍:
DAO publishes Research Articles, Reviews, and Notes, as well as Comments/Reply Comments (for details see DAO 48:161), Theme Sections and Opinion Pieces. For details consult the Guidelines for Authors. Papers may cover all forms of life - animals, plants and microorganisms - in marine, limnetic and brackish habitats. DAO''s scope includes any research focusing on diseases in aquatic organisms, specifically:
-Diseases caused by coexisting organisms, e.g. viruses, bacteria, fungi, protistans, metazoans; characterization of pathogens
-Diseases caused by abiotic factors (critical intensities of environmental properties, including pollution)-
Diseases due to internal circumstances (innate, idiopathic, genetic)-
Diseases due to proliferative disorders (neoplasms)-
Disease diagnosis, treatment and prevention-
Molecular aspects of diseases-
Nutritional disorders-
Stress and physical injuries-
Epidemiology/epizootiology-
Parasitology-
Toxicology-
Diseases of aquatic organisms affecting human health and well-being (with the focus on the aquatic organism)-
Diseases as indicators of humanity''s detrimental impact on nature-
Genomics, proteomics and metabolomics of disease-
Immunology and disease prevention-
Animal welfare-
Zoonosis