title: Real-time PCR detection of Campylobacter spp.: A comparison to classic culturing and enrichment.
abstract: The major disadvantage of the current gold standard for detection of the food pathogen Campylobacter, i.e. culturing, is the lengthy procedure. In this study we assessed the use of real-time PCR for detection of Campylobacter. To this end, 926 poultry samples, taken from transport containers and broiler caeca in The Netherlands in 2007, were subjected to three different real-time PCR detection methods: one targeting the Campylobacter jejuni hipO gene, one targeting the Campylobacter coli glyA gene, and one generically targeting Campylobacter spp. 16S rDNA sequence. The PCR results from the three different PCR protocols were compared to the work of Nauta et al. (2009) who analyzed the same set of samples collected from 62 broiler flocks by means of enrichment culturing. The results indicate that the generic 16S campylobacter PCR detection is equally reliable but much faster (4 h instead of ≥2 days) than detection by means of culturing. Moreover, PCR detection targeting the hipO and the glyA gene provide the possibility of C. jejuni and C. coli species discrimination. The generic Campylobacter spp. PCR analysis also confirmed the high incidence of Campylobacter spp. in poultry samples (∼90%) and the species specific PCR showed the simultaneous presence of C. jejuni and C. coli in ∼24% of the samples. Furthermore, the results from the three PCR analyses suggested the occurrence of alternative Campylobacter species in almost 10% of the samples. The campylobacter PCR detection methods reported here can replace traditional culturing because of being quicker and more reliable.
authors.affiliation: TNO, Microbiology & Systems Biology, Utrechtseweg 48, 3704 HE, Zeist, The Netherlands. Electronic address: paulo.deboer@tno.nl.
authors.forename: P
authors.initials: P
authors.lastname: de Boer
authors.affiliation: TNO, Microbiology & Systems Biology, Utrechtseweg 48, 3704 HE, Zeist, The Netherlands.
authors.forename: H
authors.initials: H
authors.lastname: Rahaoui
authors.affiliation: TNO, Microbiology & Systems Biology, Utrechtseweg 48, 3704 HE, Zeist, The Netherlands.
authors.forename: R J
authors.initials: RJ
authors.lastname: Leer
authors.affiliation: TNO, Microbiology & Systems Biology, Utrechtseweg 48, 3704 HE, Zeist, The Netherlands.
authors.forename: R C
authors.initials: RC
authors.lastname: Montijn
authors.affiliation: TNO, Microbiology & Systems Biology, Utrechtseweg 48, 3704 HE, Zeist, The Netherlands.
authors.forename: J M B M
authors.initials: JM
authors.lastname: van der Vossen
mesh_terms: D000818:Animals; D002167:Campylobacter / Q000235:genetics* / Q000254:growth & development* / Q000302:isolation & purification / Q000378:metabolism; D017000:Campylobacter coli / Q000235:genetics / Q000302:isolation & purification; D016123:Campylobacter jejuni / Q000235:genetics; D002432:Cecum / Q000382:microbiology; D002645:Chickens; D004269:DNA, Bacterial / Q000235:genetics; D011200:Poultry / Q000382:microbiology; D060888:Real-Time Polymerase Chain Reaction; D015203:Reproducibility of Results
chemical_list: D004269:DNA, Bacterial
keywords: Campylobacter; Detection; Poultry samples; Real-time PCR