{"title":"用羟基萘酚蓝通过环路介导等温扩增法开发出一种快速检测 Prorocentrum lima 的方法","authors":"Chao Yang, Yu Zhen, Jialin Hou, Tiezhu Mi","doi":"10.1007/s10126-024-10310-2","DOIUrl":null,"url":null,"abstract":"<div><p><i>Prorocentrum lima</i>, a widely distributed dinoflagellate known for its production of harmful biotoxins, poses a significant threat to humans, aquaculture, and marine ecosystems. As a result, the detection of this toxic alga in coastal waters has become an urgent research focus. In this study, a rapid, sensitive, and cost-effective detection method based on loop-mediated isothermal amplification (LAMP) was developed to identify <i>P. lima.</i> In this method, cell extracts of <i>P. lima</i> were diluted and used directly as templates for amplification, eliminating the need for nucleic acid purification and simplifying the detection process. Hydroxy naphthol blue (HNB) was incorporated into the reaction mix to facilitate result interpretation, enabling visual determination of the amplification outcome with the naked eye. The entire detection process, from DNA extraction to template amplification and product detection, could be completed within 80 min using a simple constant temperature-control device. This LAMP-based detection method demonstrated excellent reliability, specificity, and a low detection limit of 5.87 cells/mL for DNA crude extract. The assay offered an efficient alternative to PCR for rapid detection of <i>P. lima</i>. By streamlining the detection process and offering a visual readout, this technique holds promise for efficient and routine monitoring of harmful algal species, benefitting both research efforts and environmental management strategies.</p></div>","PeriodicalId":690,"journal":{"name":"Marine Biotechnology","volume":"26 3","pages":"475 - 487"},"PeriodicalIF":2.6000,"publicationDate":"2024-04-11","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"0","resultStr":"{\"title\":\"Development of a Rapid Detection Method to Prorocentrum lima by Loop-Mediated Isothermal Amplification with Hydroxy Naphthol Blue\",\"authors\":\"Chao Yang, Yu Zhen, Jialin Hou, Tiezhu Mi\",\"doi\":\"10.1007/s10126-024-10310-2\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"<div><p><i>Prorocentrum lima</i>, a widely distributed dinoflagellate known for its production of harmful biotoxins, poses a significant threat to humans, aquaculture, and marine ecosystems. As a result, the detection of this toxic alga in coastal waters has become an urgent research focus. In this study, a rapid, sensitive, and cost-effective detection method based on loop-mediated isothermal amplification (LAMP) was developed to identify <i>P. lima.</i> In this method, cell extracts of <i>P. lima</i> were diluted and used directly as templates for amplification, eliminating the need for nucleic acid purification and simplifying the detection process. Hydroxy naphthol blue (HNB) was incorporated into the reaction mix to facilitate result interpretation, enabling visual determination of the amplification outcome with the naked eye. The entire detection process, from DNA extraction to template amplification and product detection, could be completed within 80 min using a simple constant temperature-control device. This LAMP-based detection method demonstrated excellent reliability, specificity, and a low detection limit of 5.87 cells/mL for DNA crude extract. The assay offered an efficient alternative to PCR for rapid detection of <i>P. lima</i>. By streamlining the detection process and offering a visual readout, this technique holds promise for efficient and routine monitoring of harmful algal species, benefitting both research efforts and environmental management strategies.</p></div>\",\"PeriodicalId\":690,\"journal\":{\"name\":\"Marine Biotechnology\",\"volume\":\"26 3\",\"pages\":\"475 - 487\"},\"PeriodicalIF\":2.6000,\"publicationDate\":\"2024-04-11\",\"publicationTypes\":\"Journal Article\",\"fieldsOfStudy\":null,\"isOpenAccess\":false,\"openAccessPdf\":\"\",\"citationCount\":\"0\",\"resultStr\":null,\"platform\":\"Semanticscholar\",\"paperid\":null,\"PeriodicalName\":\"Marine Biotechnology\",\"FirstCategoryId\":\"99\",\"ListUrlMain\":\"https://link.springer.com/article/10.1007/s10126-024-10310-2\",\"RegionNum\":3,\"RegionCategory\":\"生物学\",\"ArticlePicture\":[],\"TitleCN\":null,\"AbstractTextCN\":null,\"PMCID\":null,\"EPubDate\":\"\",\"PubModel\":\"\",\"JCR\":\"Q3\",\"JCRName\":\"BIOTECHNOLOGY & APPLIED MICROBIOLOGY\",\"Score\":null,\"Total\":0}","platform":"Semanticscholar","paperid":null,"PeriodicalName":"Marine Biotechnology","FirstCategoryId":"99","ListUrlMain":"https://link.springer.com/article/10.1007/s10126-024-10310-2","RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q3","JCRName":"BIOTECHNOLOGY & APPLIED MICROBIOLOGY","Score":null,"Total":0}
引用次数: 0
摘要
Prorocentrum lima 是一种广泛分布的甲藻,以产生有害的生物毒素而闻名,对人类、水产养殖和海洋生态系统构成重大威胁。因此,检测沿海水域中的这种有毒藻类已成为一项紧迫的研究重点。本研究开发了一种基于环介导等温扩增(LAMP)技术的快速、灵敏且经济有效的检测方法来鉴定利马藻。在该方法中,利马蝇的细胞提取物稀释后直接用作扩增模板,无需进行核酸纯化,简化了检测过程。反应混合物中加入了羟基萘酚蓝(HNB)以方便结果判读,可通过肉眼目测扩增结果。从 DNA 提取到模板扩增再到产物检测,整个检测过程可在 80 分钟内通过简单的恒温装置完成。这种基于 LAMP 的检测方法具有极佳的可靠性和特异性,DNA 粗提取物的检测限低至 5.87 cells/mL。该检测方法为快速检测利马蝇提供了一种有效的 PCR 替代方法。通过简化检测过程并提供可视读数,该技术有望实现对有害藻类的高效常规监测,从而有利于研究工作和环境管理策略。
Development of a Rapid Detection Method to Prorocentrum lima by Loop-Mediated Isothermal Amplification with Hydroxy Naphthol Blue
Prorocentrum lima, a widely distributed dinoflagellate known for its production of harmful biotoxins, poses a significant threat to humans, aquaculture, and marine ecosystems. As a result, the detection of this toxic alga in coastal waters has become an urgent research focus. In this study, a rapid, sensitive, and cost-effective detection method based on loop-mediated isothermal amplification (LAMP) was developed to identify P. lima. In this method, cell extracts of P. lima were diluted and used directly as templates for amplification, eliminating the need for nucleic acid purification and simplifying the detection process. Hydroxy naphthol blue (HNB) was incorporated into the reaction mix to facilitate result interpretation, enabling visual determination of the amplification outcome with the naked eye. The entire detection process, from DNA extraction to template amplification and product detection, could be completed within 80 min using a simple constant temperature-control device. This LAMP-based detection method demonstrated excellent reliability, specificity, and a low detection limit of 5.87 cells/mL for DNA crude extract. The assay offered an efficient alternative to PCR for rapid detection of P. lima. By streamlining the detection process and offering a visual readout, this technique holds promise for efficient and routine monitoring of harmful algal species, benefitting both research efforts and environmental management strategies.
期刊介绍:
Marine Biotechnology welcomes high-quality research papers presenting novel data on the biotechnology of aquatic organisms. The journal publishes high quality papers in the areas of molecular biology, genomics, proteomics, cell biology, and biochemistry, and particularly encourages submissions of papers related to genome biology such as linkage mapping, large-scale gene discoveries, QTL analysis, physical mapping, and comparative and functional genome analysis. Papers on technological development and marine natural products should demonstrate innovation and novel applications.