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.

利马原甲藻是一种分布广泛的甲藻，以其产生有害生物毒素而闻名，对人类、水产养殖和海洋生态系统构成重大威胁。因此，沿海水域中这种有毒藻类的检测已成为紧迫的研究热点。在本研究中，开发了一种基于环介导等温扩增（LAMP）的快速、灵敏且经济高效的检测方法来鉴定利马疟原虫。该方法将利马细胞提取物稀释后直接作为扩增模板，无需进行核酸纯化，简化了检测过程。将羟基萘酚蓝 (HNB) 添加到反应混合物中以促进结果解释，从而能够用肉眼直观地确定扩增结果。使用简单的恒温控制装置，从DNA提取到模板扩增、产物检测的整个检测过程可以在80分钟内完成。这种基于 LAMP 的检测方法表现出出色的可靠性、特异性以及 DNA 粗提物的低检测限（5.87 个细胞/mL）。该测定法为快速检测利马疟原虫提供了 PCR 的有效替代方案。通过简化检测过程并提供视觉读数，该技术有望对有害藻类物种进行高效和常规监测，从而有利于研究工作和环境管理策略。