JOURNAL OF FOOD SAFETY, vol.In Press, 2015 (Journal Indexed in SCI Expanded)
The increasing industrial interest in easy, economical and reliable methods has led to the development and application of DNA-based methods for the detection of microbial pathogens in food. In the present paper, we describe the development of a cost-efficient EvaGreen-based real-time PCR technique for simultaneous and practical detection of Listeria monocytogenes (L. monocytogenes) and Escherichia coli O157:H7 (E. coli O157:H7) in the food matrix. EvaGreen-based simplex and duplex real-time PCR showed that specific PCR products were identified by melting curve analysis, and had a reproducible Tm of 77.00±0.4°C for L. monocytogenes and 85.60±0.2 for E. coli O157:H7.The absolute detection limit of Eva Greenbased simplex and duplex real-time PCR was 0.0001 ng/µl for DNA of both pathogens. The relative detection limit of the simplex and duplex technique was less than 10 cells/ml for the two pathogens.