Poultry Institute, Chinese Academy of Agricultural Sciences, Jiangsu, China.
Curr Microbiol. 2011 Dec;63(6):511-6. doi: 10.1007/s00284-011-0013-3. Epub 2011 Sep 21.
Loop-mediated isothermal amplification (LAMP) was designed for detection of Listeria monocytogenes, which is an important food-borne kind of pathogenic bacteria causing human and animal disease. The primers set for the hlyA gene consist of six primers targeting eight regions on specific gene. The LAMP assay could be performed within 40 min at 65°C in a water bath. Amplification products were visualized by calcein and manganous ion and agarose gel electrophoresis. Sensitivity of the LAMP assay for detection of L. monocytogenes in pure cultures was 2.0 CFU per reaction. The LAMP assay was 100-fold higher sensitive than that of the conventional PCR assay. Taking this way, 60 chicken samples were investigated for L. monocytogenes. The accuracy of LAMP was shown to be 100% when compared to the "gold standard" culture-biotechnical, while the PCR assay failed to detect L. monocytogenes in two of the positive samples. It is shown that LAMP assay can be used as a sensitive, rapid, and simple detection tool for the detection of L. monocytogenes and will facilitate the surveillance for contamination of L. monocytogenes in food.
环介导等温扩增（LAMP）被设计用于检测李斯特菌，它是一种重要的食源性致病菌，会导致人类和动物疾病。针对 hlyA 基因的引物集由针对特定基因八个区域的六个引物组成。LAMP 检测可以在 65°C 的水浴中 40 分钟内进行。通过钙黄绿素和锰离子以及琼脂糖凝胶电泳可视化扩增产物。LAMP 检测在纯培养物中检测李斯特菌的灵敏度为 2.0 CFU/反应。LAMP 检测比传统 PCR 检测灵敏 100 倍。通过这种方式，对 60 个鸡肉样本进行了李斯特菌检测。与“金标准”培养生物技术相比，LAMP 的准确率为 100%，而 PCR 检测未能在两个阳性样本中检测到李斯特菌。结果表明，LAMP 检测可作为一种敏感、快速、简单的检测工具，用于检测李斯特菌，并有助于监测食品中李斯特菌的污染。
