title: A multiplex polymerase chain reaction to detect and differentiate Campylobacter fetus subspecies fetus and Campylobacter fetus -species venerealis: use on UK isolates of C. fetus and other Campylobacter spp.
abstract: OBJECTIVE
  Subspeciation of Campylobacter fetus subsp. fetus (CFF) and Campylobacter fetus subsp. venerealis (CFV) is important for international animal import regulations. Phenotyping can be unreliable, and genotyping by techniques like pulsed field gel electrophoresis is difficult in routine diagnostic laboratories. A PCR subspeciation technique has been reported [Aust Vet J (1997) 75, 827]; we aimed to develop this PCR and investigate its use on UK C. fetus isolates.
  
  
  RESULTS
  We augmented the PCR with further primers, and tested 76 isolates of C. fetus and 16 isolates of other Campylobacter spp. PCR failed to correlate well with phenotyping, especially for CFV. We characterized the amplicon of the CFV-specific primers (reported as plasmid derived, but unavailable on the public databases); and predicted a parA gene sequence, anticipated to be plasmid-associated. However, although plasmid isolations from selected CFV isolates demonstrated the presence of several plasmids, there was no correlation between plasmid profile and PCR result. Further, the parA sequence was not detected by PCR in any of the plasmid bands.
  
  
  CONCLUSIONS
  This PCR is not suitable for subspeciation of C. fetus in the UK. The results suggest that this is a reflection of the presence of an unusual clone of CFV currently present in cattle in this country.
  
  
  CONCLUSIONS
  PCR cannot substitute for phenotyping of C. fetus isolates in the UK. The reasons for failure of PCR genotyping may reflect local strains and/or plasmid profiles. Further study is required to better elucidate molecular sub-speciation of C. fetus.
authors.affiliation: Moredun Research Institute, Bush Loan, Penicuik, Midlothian, UK. willk@mri.sari.ac.uk
authors.forename: K
authors.initials: K
authors.lastname: Willoughby
authors.forename: P F
authors.initials: PF
authors.lastname: Nettleton
authors.forename: M
authors.initials: M
authors.lastname: Quirie
authors.forename: M A
authors.initials: MA
authors.lastname: Maley
authors.forename: G
authors.initials: G
authors.lastname: Foster
authors.forename: M
authors.initials: M
authors.lastname: Toszeghy
authors.forename: D G
authors.initials: DG
authors.lastname: Newell
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D001426:Bacterial Proteins / Q000235:genetics; D001483:Base Sequence; D002167:Campylobacter / Q000235:genetics / Q000302:isolation & purification*; D002168:Campylobacter fetus / Q000235:genetics / Q000302:isolation & purification; D002417:Cattle; D017931:DNA Primers / Q000235:genetics; D005798:Genes, Bacterial / Q000235:genetics; D005838:Genotype; D010957:Plasmids / Q000235:genetics; D016133:Polymerase Chain Reaction / Q000379:methods*; D013045:Species Specificity; D006113:United Kingdom
chemical_list: D001426:Bacterial Proteins; D017931:DNA Primers; C052883:chromosome partition proteins, bacterial