环介导等恒扩增技术(LAMP)是一种特异、灵敏、快速的新型基因检测技术。实验以单增李斯特菌为研究对象,设计了4种LAMP引物以特异地识别单增李斯特菌中gyrB基因的6个特殊区域,并进行LAMP反应,建立单增李斯特菌的LAMP快速检测技术。结果表明,LAMP技术能在恒温条件下,1h内检测出单增李斯特菌,灵敏度达到6.4×101CFU/reaction,并且与其他常见的细菌无交叉反应。其对人工感染牛奶样品的检出量为1CFU/mL,适合于食品中污染单增李斯特菌的快速检测。

Loop-mediated isothermal amplification(LAMP) method is a novel gene detection technique with specificity,sensitive and rapidity.In this paper,we developed and evaluated the specificity and sensitivity of the LAMP assay for rapid detection of Listeria monocytogenes.A set of four LAMP primers were designed for recognizing six distinct regions on the target DNA(gyrB).The results showed that the LAMP assay was able to specifically detect all of the L.monocytogenes strains without amplification from other strains.The detection limit was 6.4×10-1CFU/reaction in pure culture.When applied in spiked oysters,the detection limit was 1 CFU/mL.So the LAMP method is good for detection of L.monocytogenes for food safety.