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Gut Pathogens
Published in: Gut Pathogens 1/2014

Open Access 01-12-2014 | Research

Detection, identification and quantification of Campylobacter jejuni, coli and lari in food matrices all at once using multiplex qPCR

Authors: Lucie Vondrakova, Jarmila Pazlarova, Katerina Demnerova

Published in: Gut Pathogens | Issue 1/2014

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Abstract

Background

Thermotolerant Campylobacter jejuni, coli and lari are recognized as leading food-borne pathogens causing an acute bacterial enteritis worldwide. Due to narrow spectrum of their biochemical activity, it is very complicated to distinguish between individual species. For reliable risk assessment, proper incidence evaluation or swift sample analysis regarding individual species, a demand for simple and rapid method for their distinguishing is reasonable. In this study, we evaluated a reliable and simple approach for their simultaneous detection, species identification and quantification using multiplex qPCR.

Results

Species specific primers and hydrolysis probes are directed to hippuricase gene of C. jejuni, serine hydroxymethyltransferase gene of C. coli and peptidase T gene of C. lari. Efficiencies of reactions were 90.85% for C. jejuni, 96.97% for C. coli and 92.89% for C. lari. At 95.00% confidence level and when cut off is set to 38 cycles, limits of detection are in all cases under 10 genome copies per reaction which is very appreciated since it is known that infectious doses are very low.

Conclusions

Proposed assay was positively validated on different food matrices (chicken wing rinses, chicken juice and homogenized fried chicken strips). No inhibition of PCR reaction occurred. Assay was evaluated in accordance with MIQE handbook.
Appendix
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Metadata
Title
Detection, identification and quantification of Campylobacter jejuni, coli and lari in food matrices all at once using multiplex qPCR
Authors
Lucie Vondrakova
Jarmila Pazlarova
Katerina Demnerova
Publication date
01-12-2014
Publisher
BioMed Central
Published in
Gut Pathogens / Issue 1/2014
Electronic ISSN: 1757-4749
DOI
https://doi.org/10.1186/1757-4749-6-12

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