环介导等温扩增法检测动植物源性单核细胞增生李斯特氏菌

Detection of Listeria monocytogenes in animal and plant derived food by loop-mediated isothermal amplification

中文关键词:  环介导等温扩增  单核细胞增生李斯特氏菌  动植物源性

英文关键词:loop-mediated isothermal amplification  Listeria monocytogenes  animal and plant origin

中文摘要:

      目的 采用环介导等温扩增技术检测动植物源性单核细胞增生李斯特氏菌。方法 在北京市范围内各业态餐厅购买600份凉菜样品为研究对象, 以单核细胞增生李斯特氏菌的hlyA基因为目标基因, 设计4条特异性引物, 使用环比等温扩增法(loop-to-isothermal amplification, LAMP)创建凉菜中单核细胞增生李斯特氏菌快速检测方法并将其应用于鉴定凉菜中分离的单核细胞增生李斯特氏菌。结果 通过细菌分离培养法对600份凉菜进行检测, 一共检测分离出37株致病菌菌株, 其中包括7株单核细胞增生李斯特氏菌、10株沙门氏菌和20株大肠埃希氏菌, 单核细胞增生李斯特氏菌的检出率为1.2%。用LAMP对样品中的菌株进行测定, 7株凉菜中分离的单核细胞增生李斯特氏菌为阳性, 其他菌株为阴性。通过LAMP方法检测动植物源性凉菜中分离的单核细胞增生李斯特氏菌平均检出限为2.2*10 CFU/μL、特异性为100%。动物源性凉菜中的致病菌含量较高。结论 环介导等温扩增法具备灵敏、快速、操作简便的特点, 适合用于餐饮行业的现场快速检测领域。

英文摘要:

      Objective To detect Listeria monocytogenes of animal and plant origin by loop-mediated isothermal amplification. Methods Total of 600 cold dish samples were purchased from restaurants of various formats in Beijing. Taking the hlyA gene of Listeria monocytogenes as the target gene, 4 specific primers were designed, and the Listeria monocytogenes in cold dishes was rapidly detected using loop-to-isothermal amplification (LAMP), which was also used to identify Listeria monocytogenes isolated in cold dishes. Results A total of 600 cold dishes were tested by bacterial isolation and culture, and 37 strains of pathogenic bacteria were detected, including 7 strains of Listeria monocytogenes, 10 strains of Salmonella, and 20 strains of Escherichia coli. The detection rate of Listeria monocytogenes was 1.2%. LAMP was used to determine the strains in the samples. Seven strains of Listeria monocytogenes isolated from the cold dishes were positive, and other strains were negative. The average detection limit of Listeria monocytogenes in cold dishes of animal and plant origin by the LAMP method was 2.2*10 CFU/μL and the specificity was 100%. The content of pathogenic bacteria in cold dishes of animal origin was high. Conclusion The loop-mediated isothermal amplification method is sensitive, fast, and easy to operate, which is suitable for on-site rapid detection in the restaurant industry.