Original Scientific Article
Development of simple multiplex real-time PCR assays for foodborne pathogens detection and identification on LightCycler
Avo Karus * ,
Fabrizio Ceciliani ,
Armand Sanches Bonastre ,
Virge Karus

Mac Vet Rev 2017; 40 (1): 53 - 58

10.1515/macvetrev-2017-0010

Received: 18 November 2016

Received in revised form: 22 December 2016

Accepted: 24 December 2016

Available Online First: 03 January 2017

Published on: 15 March 2017

Correspondence: Avo Karus, avo.karus@emu.ee
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Abstract

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.

Keywords: food-borne pathogens, multiplex real-time PCR, melting curve genotyping


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Copyright

© 2017 Karus A. This is an open-access article publishedunder the terms of the Creative Commons Attribution License whichpermits unrestricted use, distribution, and reproduction in any medium,provided the original author and source are credited.

Conflict of Interest Statement

The authors declared that they have no potential conflict of interest with respect to the authorship and/or publication of this article.

Citation Information

Macedonian Veterinary Review. Volume 40, Issue 1, Pages 53-58, p-ISSN 1409-7621, e-ISSN 1857-7415, DOI: 10.1515/macvetrev-2017-0010, 2017