ORIGINAL_ARTICLE 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. https://macvetrev.mk/Files/Article/2019/10.1515/macvetrev-2017-0010/macvetrev-2017-0010.pdf 2017-03-15T09:00:00 53 58 10.2478/macvetrev-2017-0010 food-borne pathogens multiplex real-time PCR melting curve genotyping Avo Karus avo.karus@emu.ee false 1 Department of Food Sciences and Food Technology, Institute of Veterinary Medicine and Animal Science, Estonian University of Life Sciences, Kreutzwaldi 62, Tartu, Estonia LEAD_AUTHOR Fabrizio Ceciliani false 2 Department of Veterinary Science and Public Health, University of Milan, via Celoria 10, 20133 Milan, Italy AUTHOR Armand Sanches Bonastre false 3 Universitat Autonoma de Barcelona, Campus de la UAB, Bellaterra, Spain AUTHOR Virge Karus false 4 Department of Food Sciences and Food Technology, Institute of Veterinary Medicine and Animal Science, Estonian University of Life Sciences, Kreutzwaldi 62, Tartu, Estonia AUTHOR Fleckenstein JM, Bartels SR, Drevets PD, Bronze MS, Drevets DA, Infectious agents of food- and water-borne illnessesAm J Med Sci 2010; 340: 3238-246. 1 10.1097/MAJ.0b013e3181e99893 Postollec F, Falentin H, Pavan S, Combrisson J, Sohier D, Recent advances in quantitative PCR (qPCR) applications in food microbiologyFood Microbiol 2011; 28: 5848-861. 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