Development of Simple Multiplex Real-Time PCR Assays for Foodborne Pathogens Detection and Identification On Lightcycler

Avo Karus 1 , Fabrizio Ceciliani 2 , Armand Sanches Bonastre 3 , und Virge Karus 1
  • 1 Department of Food Sciences and Food Technology, Institute of Veterinary Medicine and Animal Science, Estonian University of Life Sciences, Tartu, Estonia
  • 2 Department of Veterinary Science and Public Health, University of Milan, Milan, Italy
  • 3 Universitat Autonoma de Barcelona, , Spain


Most acute intestinal diseases are caused by food-borne pathogens. A fast and simple real-time PCR-based procedure for simultaneous detection of food contamination by any of the five food-borne pathogens: Campylobacter jejuni, Mycobacterium bovis, Enterobacter sakazaki, Shigella boydii, Clostridium perfrigens using multiplex EvaGreen real-time PCR for LightCycler was developed and evaluated. Real-time qPCR showed excellent sensitivity. Tm calling and Melting Curve Genotyping (MCG) were used for analysis of PCR product melting curves. The Melting Curve Genotyping option showed good performance for discrimination of positive samples containing DNA of single pathogen or pathogen mixtures from negative samples.

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