Development and application of a loop-mediated isothermal amplification method on rapid detection Escherichia coli O157 strains from food samples

被引:162
作者
Zhao, Xihong [1 ]
Li, Yanmei [2 ]
Wang, Li [3 ]
You, Lijun [1 ]
Xu, Zhenbo [1 ,4 ]
Li, Lin [1 ]
He, Xiaowei [1 ]
Liu, Yao [5 ]
Wang, Jihua [6 ]
Yang, Liansheng [1 ]
机构
[1] S China Univ Technol, Coll Light Ind & Food Sci, Guangzhou 510640, Guangdong, Peoples R China
[2] Guangzhou Med Univ, Guangzhou, Guangdong, Peoples R China
[3] S China Agr Univ, Coll Food Sci, Food Safety Key Lab Guangdong Prov, Guangzhou, Guangdong, Peoples R China
[4] Univ Maryland Baltimore, Sch Dent, Dept Microbial Pathogenesis, Baltimore, MD 21201 USA
[5] Zhongshan Supervis Testing Inst Qual & Metrol, Zhongshan, Peoples R China
[6] Guangzhou Wondfo Biotechnol Co, Guangzhou, Guangdong, Peoples R China
基金
国家教育部科学基金资助; 中国国家自然科学基金;
关键词
Loop-mediated isothermal amplification (LAMP); Escherichia coli O157; Rapid detection; SALMONELLA; CULTURE; CHINA;
D O I
10.1007/s11033-009-9700-6
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
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.
引用
收藏
页码:2183 / 2188
页数:6
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