作　　者： ; ; ; ; ; ;

机构地区： 河北农业大学食品科技学院

出　　处： 《中国食品学报》 2012年第4期168-174,共7页

摘　　要： 目的:建立环介导等温扩增技术(LAMP)快速检测产气荚膜梭菌的方法。方法:以产气荚膜梭菌(ATCC13124)的α毒素全基因序列为保守序列,设计内、外、环引物,肉眼观察白色沉淀并判断检测结果。结果:LAMP检测产气荚膜梭菌的灵敏度为2.92×102CFU/mL,人工污染产气荚膜梭菌的全脂乳的检测限为15.7CFU/mL,耗时仅1h。对照PCR检测产气荚膜梭菌的灵敏度为2.92×105CFU/mL,人工污染产气荚膜梭菌的全脂乳的检出限1.57×105CFU/mL。采用同样的方法提取DNA,耗时3h。结论:建立LAMP快速、特异检测产气荚膜梭菌的方法,该方法灵敏度是PCR的1000倍,为食品中产气荚膜梭菌的检测构建了一个技术平台。 Objective： An assay using loop-mediated isothermal amplification was developed for rapidly detection of Clostridium perfringens. Method： Sequences of alpha-toxin of Clostridium perfrigens（ATCC13124） were used as target sequences to design outer primers, inner primers and loop primers. We judge the results through the visible by naked eye of the white precipitate. Results： The sensitivity of the LAMP assay was 2.92×10^2 CFU/mL and the detection limit of artificial contamination was 15.7 CFU/mL, the detection could be finished in an hour. Compared with LAMP, the sensitivity of the PCR assay was 2.92×10^5 CFU/mL and the detection limit of artificial contamination was 1.57×10^5 CFU/mL. When we used the same method to extract DNA, it would be spend three hours. Conclusion： A rapid and special method was established for detection of Clostridium pe（frigens, the sensitivity of the LAMP were 1 000 times as high as that of PCR. This result indicates that LAMP can provide a technical platform for rapid detection of Clostridium pe^rfigens in food.

关 键 词： 环介导等温扩增 检测 产气荚膜梭菌 毒素

领　　域： [农业科学] [农业科学] [农业科学]