Loop-mediated isothermal amplification for detecting enterohemorrhagic Escherichia coli O157:H7: a comparison with PCR.
- Author:
Rong JIANG
1
;
Beiguo LONG
;
Guifen ZENG
;
Dan WANG
;
Hongying FAN
;
Xianbo WU
Author Information
- Publication Type:Journal Article
- MeSH: DNA, Bacterial; Escherichia coli O157; isolation & purification; Nucleic Acid Amplification Techniques; methods; Polymerase Chain Reaction; methods; Sensitivity and Specificity
- From: Journal of Southern Medical University 2012;32(7):1026-1030
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVETo establish a rapid method of loop-mediated isothermal amplification (LAMP) for detecting enterohemorrhagic Escherichia coli (EHEC) O157:H7.
METHODSSix primers that specifically recognized the rfbE gene of EHEC O157:H7 were designed. Under the optimized reaction conditions, LAMP and PCR were evaluated for the sensitivity and specificity in the detection of 39 laboratory samples of EHEC O157:H7 strains, and their detection results of contaminated fresh pork samples were compared.
RESULTSLAMP assay correctly identified all the 7 EHEC O157:H7 strains and showed negative results for all the 32 non-EHEC O157:H7 strains. The detection limit of LAMP was much lower than that of rfbE-PCR (10 vs 100 cfu/ml). In the detection of the contaminated pork samples, both LAMP and PCR yielded results consistent with those by the conventional detection method.
CONCLUSIONThe rfbE-based LAMP assay can serve as a rapid, sensitive, specific and low-cost means for detecting EHEC O157:H7 strain.
