The detection and monitoring of Vibrio parahaemolyticus pathogen in aquatic foods have become essential for preventing outbreaks. In this study, loop-mediated isothermal amplification (LAMP) assay with the azo dye, hydroxynaphthol blue (HNB) was developed targeting species-specific tlh gene. The assay was carried out on 62 seafood samples that included clam and shrimp and compared with conventional LAMP assay performed with the commonly used fluorescent dye, conventional PCR, and real-time PCR (RT-PCR). Of 62 samples studied for tlh gene, 32 (51.61%) gave positive by HNB-LAMP, which comprised 22 (70.96%) clam samples and 10 (32.25%) shrimp samples. The HNB-LAMP assay was found to be highly sensitive, specific, and superior to conventional PCR (p > 0.05). RT-PCR presented higher sensitivity than HNB-LAMP; however, it has the limitation of being cost-intensive and requiring technical expertise to perform. HNB-LAMP is affordable, rapid, simple, and easy to perform, allowing naked eye visualization.

中文翻译：

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通过环介导等温扩增和羟基萘酚蓝染料快速目测海鲜样品中的副溶血性弧菌

水产食品中副溶血性弧菌病原体的检测和监测对于预防爆发至关重要。在本研究中，针对物种特异性 tlh 基因开发了使用偶氮染料羟基萘酚蓝 (HNB) 的环介导等温扩增 (LAMP) 检测。该测定对包括蛤和虾在内的 62 个海鲜样品进行，并与使用常用荧光染料、常规 PCR 和实时 PCR (RT-PCR) 进行的常规 LAMP 测定进行比较。在研究 tlh 基因的 62 个样本中，32 个（51.61%）HNB-LAMP 呈阳性，其中包括 22 个（70.96%）蛤蜊样本和 10 个（32.25%）虾样本。发现 HNB-LAMP 检测具有高度灵敏性、特异性且优于常规 PCR (p > 0.05)。RT-PCR 的灵敏度高于 HNB-LAMP；然而，它具有成本密集型和需要技术专长才能执行的限制。HNB-LAMP 价格实惠、快速、简单且易于执行，可实现肉眼可视化。