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.

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