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

Abstract:

Article Preview

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.

Info:

Periodical:

Advanced Materials Research (Volumes 343-344)

Edited by:

David Wang

Pages:

1217-1221

DOI:

10.4028/www.scientific.net/AMR.343-344.1217

Citation:

D. G. Wang and G. C. Huo, "Rapid Detection Viable Escherichia Coli O157 in Raw Milk Using Loop-Mediated Isothermal Amplification with Aid of Ethidium Monoazide", Advanced Materials Research, Vols. 343-344, pp. 1217-1221, 2012

Online since:

September 2011

Export:

Price:

$35.00

In order to see related information, you need to Login.

In order to see related information, you need to Login.