Development and application of a loop-mediated isothermal amplification method on rapid detection Escherichia coli O157 strains from food samples.
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.
Zhao
Xihong
X
College of Light Industry and Food Sciences, South China University of Technology, 510640 Guangzhou, China.
Li
Yanmei
Y
Wang
Li
L
You
Lijun
L
Xu
Zhenbo
Z
Li
Lin
L
He
Xiaowei
X
Liu
Yao
Y
Wang
Jihua
J
Yang
Liansheng
L
D001681:Biological Assay; D017931:DNA Primers / Q000378:metabolism; D019453:Escherichia coli O157 / Q000235:genetics / Q000302:isolation & purification*; D005504:Food Analysis / Q000379:methods*; D005516:Food Microbiology; D016133:Polymerase Chain Reaction / Q000379:methods*; D012015:Reference Standards; D012680:Sensitivity and Specificity; D013696:Temperature
D017931:DNA Primers
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