基于DNA环介导等温扩增技术快速检测溶藻弧菌的研究

溶藻弧菌是引起海产鱼类、虾、贝等细菌性疾病的主要病原之一,严重危害水产养殖业的发展,建立快速、准确的检测方法对防控溶藻弧菌及保障水产养殖安全具有重要意义。本研究引入环介导等温扩增(loop-mediated isothermal amplification, LAMP)技术,以溶藻弧菌胶原酶基因为靶基因,建立了溶藻弧菌LAMP快速检测方法,45~60 min内即可完成检测。试验结果显示,该LAMP方法可特异性检测溶藻弧菌,且灵敏度高,对纯培养溶藻弧菌的检测灵敏度约10 CFU/mL,对污染水产品中溶藻弧菌的检测灵敏度为19 CFU/mL。本研究建立的溶藻弧菌LAMP检测方法操作简便、检测快速,具有良好的实用性。

Rapid Detection of Vibrio alginolyticus by Using Loop-mediated Isothermal Amplification Assay

Vibrio alginolyticus was one of the main pathogens that caused bacterial diseases in fish, shrimp and cowry, and seriously caused harm to the development of aquaculture. Thus, the development of a method to rapidly and accurately detect V.alginolyticus and protect the development of aquaculture was of great importance. In this work, a loop-mediated isothermal amplification (LAMP) method for the rapid detection of V.alginolyticus was developed using the collagenase gene of V.alginolyticus as target gene, and the LAMP test could be completed in 45 to 60 min. Results showed that the LAMP method showed a high specificity to V.alginolyticus and a good sensitivity that 10 CFU/mL for cultivated V.alginolyticus and 19 CFU/mL for its contaminated food. The LAMP method developed in this work was a sensitive, rapid and simple tool for the detection of V.alginolyticus.