title: Detection, identification and quantification of Campylobacter jejuni, coli and lari in food matrices all at once using multiplex qPCR.
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.
authors: Institute of Chemical Technology, Prague, Faculty of Food and Biochemical Technology, Department of Biochemistry and Microbiology, Technicka 5, Prague 166 28, Czech Republic.
authors: Lucie
authors: L
authors: Vondrakova
authors: Institute of Chemical Technology, Prague, Faculty of Food and Biochemical Technology, Department of Biochemistry and Microbiology, Technicka 5, Prague 166 28, Czech Republic.
authors: Jarmila
authors: J
authors: Pazlarova
authors: Institute of Chemical Technology, Prague, Faculty of Food and Biochemical Technology, Department of Biochemistry and Microbiology, Technicka 5, Prague 166 28, Czech Republic.
authors: Katerina
authors: K
authors: Demnerova
keywords: MIQE; Multiplex qPCR; Quantification; Thermotolerant Campylobacter spp