环境水体中沙门菌环介导等温扩增快速检测方法

Detection of Salmonella in water bodies by LAMP rapid method

目的建立环境水体中沙门菌的环介导等温扩增(loop-mediated isothermal amplification,LAMP)快速检测方法。方法以沙门菌的特异性基因(phoP)序列为靶序列,于63℃恒温条件下扩增1 h,80℃灭活10 min,扩增产物通过肉眼观察即可判断检测结果。结果该方法对超纯水加标水样中沙门菌的检测限可达3.07 fg/μl,对环境加标水样的检出限为2.39×10 cfu/ml。结论该方法灵敏度高、耗时短、方法简便,适用于现场或者基层实验室进行环境水体中病原微生物的快速检测。

Objective A loop-mediated isothermal amplification （LAMP） method was established as substitute for time- consuming traditional detection of Salmonella spp. in environmental water body. Methods Primers for detecting Salmonella spp. were designed based on the phoP gene of Salmonella spp. The reaction conditions were verified at 63 ℃ for 60 rain and then at 80 ℃ for 10 min,and the detection results were judged by naked eye. Results The sensitivity of the LAMP assay was found to be 3.07 fg/μl for the genome of cultivated Salmonella,and 2.39×10 cfu/ml for simulated contaminated environmental water samples. Conclusion LAMP assay is very rapid,sensitive and convenient for the detection of Salmonella spp. in environmental water, and it can be an alternative choice for the rapid detection of water-borne pathogens.