摘要目的 将环介导等温扩增(loop-mediated isothermal amplification,LAMP)方法应用于食品中单核细胞增生李斯特菌的检验,并在检测方法特异性、灵敏度等方面与实时荧光PCR和传统方法进行比较.方法 针对单核细胞增生李斯特菌hly溶血素基因设计LAMP引物,应用LAMP方法对88株单核细胞增生李斯特菌、1株单核细胞增生李斯特菌参考菌株ATCC 15313、33株非目标菌进行检测；并进行食品基质添加实验及对样品进行检测,同时应用实时荧光PCR方法和ISO 11290-1方法进行平行检测.对3种检测方法的特异性、灵敏度、检测低限及实际样品检验的结果进行比较.结果 LAMP和荧光PCR对89株单核细胞增生李斯特菌的检验结果均为阳性(100％,89/89),33株非目标菌的检测结果均为阴性(100％,33/33).LAMP方法的检测灵敏度为2×102 CFU/ml,与实时荧光PCR方法(2×102 CFU/ml)相同,且优于ISO 11290-1方法(2×104 CFU/ml).食品基质添加实验结果显示,3种检测方法的检测低限均为3 CFU/25 g样品.实际食品样品检测结果显示,3种方法单核细胞增生李斯特菌的检出率均为4％(2/45).结论 本研究建立的单核细胞增生李斯特菌LAMP检测方法具有良好的特异性,检测灵敏度与实时荧光PCR相当,适用于单核细胞增生李斯特菌的快速筛选.

abstractsObjective The loop-mediated isothermal amplification (LAMP) detection method was applied to detect Listeria monocytogenes in food.The specificity and sensitivity of this method were evaluated through comparing it with Real-time PCR and conventional detection method.Methods The LAMP primers were designed based on hly gene of Listeria monocytogenes.The LAMP method was applied to detect 88 Listeria monocytogenes,1 reference strain ATCC 15313 of Listeria monocytogenes and 33 non-targets bacteria strains; base-material addition test and practical food samples detection were also conducted.Both of Realtime PCR and ISO 11290-1 methods were used as parallel detection method in addition to LAMP.The three kinds of methods were compared by specificity,sensitivity,detection limit and the detection result of practical food samples.Results Both detection results of LAMP and Real-time PCR for 89 Listeria monocytogenes were positive (100％,89/89),33 non-targets bacteria strains were negative (100％,33/33).The sensitivity of LAMP was 2 × 102 CFU/ml,which was consistent with Real-time PCR method (2 × 102 CFU/ml) and better than ISO 11290-1 method (2 × 104 CFU/ml).Base-material addition test result showed that the detection limit of the three kinds of methods were 3 CFU/25 g samples.And the result of practical food samples displayed the same detection rate of 4％ in the three methods (2/45).Conclusion The LAMP method of Listeria monocytogenes established in this study has good specificity and sensitivity,which can be applied to the rapid detection of Listeria monocytogenes.