A highly specific and sensitive loop-mediated isothermal amplification method for the detection of Escherichia coli O157:H7

2016 ◽  
Vol 91 ◽  
pp. 161-165 ◽  
Author(s):  
Hadi Ravan ◽  
Mojdeh Amandadi ◽  
Nima Sanadgol
Keyword(s):  
Escherichia Coli ◽  
Isothermal Amplification ◽  
Escherichia Coli O157 ◽  
Loop Mediated Isothermal Amplification ◽  
Amplification Method

scholarly journals Visual Detection of Enterohemorrhagic Escherichia coli O157:H7 Using Loop-Mediated Isothermal Amplification

10.19082/2576 ◽  
2016 ◽  
Vol 8 (6) ◽  
pp. 2576-2585 ◽  
Author(s):  
Reza Ranjbar ◽  
Maryam Erfanmanesh ◽  
Davoud Afshar ◽  
Mohsen Mohammadi ◽  
Omar Ghaderi ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Visual Detection ◽  
Enterohemorrhagic Escherichia Coli ◽  
Isothermal Amplification ◽  
Escherichia Coli O157 ◽  
Loop Mediated Isothermal Amplification

Rapid Detection Viable Escherichia Coli O157 in Raw Milk Using Loop-Mediated Isothermal Amplification with Aid of Ethidium Monoazide

2011 ◽  
Vol 343-344 ◽  
pp. 1217-1221 ◽  
Author(s):  
De Guo Wang ◽  
Gui Cheng Huo
Keyword(s):  
Escherichia Coli ◽  
Thermal Processing ◽  
Lamp Assay ◽  
Raw Milk ◽  
Isothermal Amplification ◽  
Escherichia Coli O157 ◽  
Detection Level ◽  
Ethidium Monoazide ◽  
Loop Mediated Isothermal Amplification ◽  
Promising Application

The distinction between viable and dead cells was a major issue in detection of pathogenic microbe in foods especially when foods had been subjected to thermal processing. The performance of a loop-mediated isothermal amplification (LAMP) assay with aid of ethidium monoazide (EMA) for detecting viable Escherichia coli O157 in raw milk was presented in this paper. Three pairs of primers were specially designed for recognizing eight distinct sequences of rfbE gene. LAMP can only amplified DNA of viable Escherichia coli O157 because EMA selectively penetrated dead cells and covalently bound to DNA, detection limit level for artificially contaminated raw milk samples by the EMA-LAMP assay was 440 cfu/mL corresponding to 3–5 cells per reaction tube, while the detection level by EMA-PCR was 4.4×104 cfu/mL. In conclusion, EMA-LAMP had offered a novel assay for distinction between viable and dead cells with promising application in food safety detection.

Detection of Enteroaggregative Escherichia coli by Loop-Mediated Isothermal Amplification

2010 ◽  
Vol 73 (6) ◽  
pp. 1064-1072 ◽  
Author(s):  
EIJI YOKOYAMA ◽  
MASAKO UCHIMURA ◽  
KENITIRO ITO
Keyword(s):  
Escherichia Coli ◽  
Food Samples ◽  
Isothermal Amplification ◽  
Lamp Method ◽  
Bacterial Strains ◽  
Exact Test ◽  
Loop Mediated Isothermal Amplification ◽  
Amplification Method ◽  
Contaminated Food ◽  
Better Than

A novel gene amplification method, loop-mediated isothermal amplification (LAMP), has been recently developed as a rapid, specific diagnostic method for various infectious diseases. We have investigated whether LAMP can be used to detect small numbers of enteroaggregative Escherichia coli (EAEC) cells contaminated in food samples. Primers for LAMP reaction were designed with EAEC aggR gene sequences (available in GenBank). LAMP specificity with these primers was the same as that of PCR in a study of 37 EAEC and 42 non-EAEC bacterial strains. The sensitivity of the LAMP method was better than that of PCR in a study of serially diluted EAEC cells. The LAMP method was significantly more effective than was PCR in detecting EAEC-contaminated food samples (Fisher's exact test, P < 0.05). Therefore, the LAMP method described here should be useful for detecting small numbers of EAEC cells in food samples.

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