Development and application of a loop-mediated isothermal amplification method on rapid detection Escherichia coli O157 strains from food samples |
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| Authors: | Xihong Zhao Yanmei Li Li Wang Lijun You Zhenbo Xu Lin Li Xiaowei He Yao Liu Jihua Wang Liansheng Yang |
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| Affiliation: | (1) College of Light Industry and Food Sciences, South China University of Technology, 510640 Guangzhou, China;(2) Guangzhou Medical College, Guangzhou, China;(3) Food Safety Key Laboratory of Guangdong Province, College of Food Science, South China Agricultural University, Guangzhou, China;(4) Department of Microbial Pathogenesis, Dental School, University of Maryland-Baltimore, Baltimore, MD 21201, USA;(5) Zhongshan Supervision Testing Institute of Quality & Metrology, Zhongshan, China;(6) Guangzhou Wondfo Biotechnology Company, Guangzhou, China; |
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| Abstract: | We developed and evaluated the specificity and sensitivity of a loop-mediated isothermal amplification (LAMP) method for rapid detection of the food-borne Escherichia coli O157 strains. Six primers, including outer primers, inner primers and loop primers, were specially designed for recognizing eight distinct sequences on three targets, which were rfbE, stx1 and stx2. The detection limits were found to be 100, 100 and 10 fg DNA/tube for rfbE, stx1 and stx2, respectively. Application of LAMP assays were performed on 417 food-borne E. coli strains, the sensitivity of LAMP assays for the rfbE, stx1 and stx2 was 100, 95.3 and 96.3%, and the negative predictive value was 100, 96.7 and 97.1%, respectively; with a 100% specificity and positive predictive value for all three targets. |
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