A rapid and cost-efficient technique for simultaneous/duplex detection of Listeria monocytogenes and Escherichia coli O157:H7 using real time PCR

KAYNAK A., ŞAKALAR E.

JOURNAL OF FOOD SAFETY, vol.In Press, 2015 (SCI-Expanded)

  • Publication Type: Article / Article
  • Volume: In Press
  • Publication Date: 2015
  • Journal Name: JOURNAL OF FOOD SAFETY
  • Journal Indexes: Science Citation Index Expanded (SCI-EXPANDED), Scopus
  • Çanakkale Onsekiz Mart University Affiliated: Yes

Abstract

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.