Development of real-time PCR and hybridization methods for detection and identification of thermophilic Campylobacter spp. in pig faecal samples.
OBJECTIVE
To develop a real-time (rt) PCR for species differentiation of thermophilic Campylobacter and to develop a method for assessing co-colonization of pigs by Campylobacter spp.


RESULTS
The specificity of a developed 5' nuclease rt-PCR for species-specific identification of Campylobacter jejuni, Campylobacter coli, Campylobacter lari, Campylobacter upsaliensis and of a hipO gene nucleotide probe for detection of C. jejuni by colony-blot hybridization were determined by testing a total of 75 reference strains of Campylobacter spp. and related organisms. The rt-PCR method allowed species-specific detection of Campylobacter spp. in naturally infected pig faecal samples after an enrichment step, whereas the hybridization approach enhanced the specific isolation of C. jejuni (present in minority to C. coli) from pigs.


CONCLUSIONS
The rt-PCR was specific for Campylobacter jejuni, C. coli, C. lari, and C. upsaliensis and the colony-blot hybridization approach provided an effective tool for isolation of C. jejuni from pig faecal samples typically dominated by C. coli.


CONCLUSIONS
Species differentiation between thermophilic Campylobacter is difficult by phenotypic methods and the developed rt-PCR provides an easy and fast method for such differentiation. Detection of C. jejuni by colony hybridization may increase the isolation rate of this species from pig faeces.
Jensen
A N
AN
Department of Microbiological Food Safety, Danish Institute for Food and Veterinary Research, The Royal Veterinary and Agricultural University, Copenhagen, Denmark. anj@dfvf.dk
Andersen
M T
MT
Dalsgaard
A
A
Baggesen
D L
DL
Nielsen
E M
EM
D000818:Animals; D002167:Campylobacter / Q000235:genetics / Q000302:isolation & purification*; D017000:Campylobacter coli / Q000235:genetics / Q000302:isolation & purification; D016123:Campylobacter jejuni / Q000235:genetics / Q000302:isolation & purification; D044863:Campylobacter lari / Q000235:genetics / Q000302:isolation & purification; D044885:Campylobacter upsaliensis / Q000235:genetics / Q000302:isolation & purification; D015342:DNA Probes / Q000235:genetics; D004269:DNA, Bacterial / Q000235:genetics; D005243:Feces / Q000382:microbiology*; D005798:Genes, Bacterial / Q000235:genetics; D009693:Nucleic Acid Hybridization / Q000379:methods*; D016133:Polymerase Chain Reaction / Q000379:methods*; D013045:Species Specificity; D013552:Swine
D015342:DNA Probes; D004269:DNA, Bacterial