title: Detection and quantification of Campylobacter jejuni and Campylobacter coli using real-time multiplex PCR.
abstract: OBJECTIVE
  We describe a real-time quantitative multiplex polymerase chain reaction (qmPCR) assay to identify and discriminate between isolates of Campylobacter jejuni and Campylobacter coli.
  
  
  RESULTS
  Two novel sets of primers and hydrolysis probes were designed to amplify the unique DNA sequences within the hipO, ccoN and cadF genes that are specific to Camp. jejuni and Camp. coli. Using the designed optimized qmPCR assay conditions, the amplification efficiency is in range from 108 to 116%. These qmPCR assays are highly specific for Camp. jejuni and Camp. coli, as seen through testing of 40 Campylobacter strains and 17 non-Campylobacter strains. In chicken juice and tap water models spiked with known quantities of Camp. jejuni, qmPCR detected 10(2) -10(3) CFU ml(-1) within 4 h.
  
  
  CONCLUSIONS
  The qmPCR assays developed in this study provide reliable and simultaneous detection and quantification of Camp. jejuni and Camp. coli, with good amplification reaction parameters.
  
  
  CONCLUSIONS
  Following further validation, the qmPCR assay reported here has the potential to be applied to various sample types as an alternative and rapid methodology.
authors.affiliation: Omega d.o.o., Ljubljana, Slovenia. natasa.toplak@omega.si
authors.forename: N
authors.initials: N
authors.lastname: Toplak
authors.forename: M
authors.initials: M
authors.lastname: Kovač
authors.forename: S
authors.initials: S
authors.lastname: Piskernik
authors.forename: S Smole
authors.initials: SS
authors.lastname: Možina
authors.forename: B
authors.initials: B
authors.lastname: Jeršek
mesh_terms: D000818:Animals; D002169:Campylobacter Infections / Q000235:genetics; D017000:Campylobacter coli / Q000235:genetics / Q000302:isolation & purification*; D016123:Campylobacter jejuni / Q000235:genetics / Q000302:isolation & purification*; D002645:Chickens; D004269:DNA, Bacterial / Q000235:genetics; D008460:Meat / Q000382:microbiology; D060885:Multiplex Polymerase Chain Reaction / Q000379:methods*
chemical_list: D004269:DNA, Bacterial