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Background   Campylobacter  spp., especially  Campylobacter jejuni  ( C. jejuni ) and  Campylobacter coli  ( C. coli ), are recognized as the leading human foodborne pathogens in developed countries. Livestock animals carrying  Campylobacter  pose an important risk for human contamination. Pigs are known to be frequently colonized with  Campylobacter , especially  C. coli , and to excrete high numbers of this pathogen in their faeces. Molecular tools, notably real-time PCR, provide an effective, rapid, and sensitive alternative to culture-based methods for the detection of  C. coli  and  C. jejuni  in various substrates. In order to serve as a diagnostic tool supporting  Campylobacter  epidemiology, we developed a quantitative real-time PCR method for species-specific detection and quantification of  C. coli  and  C. jejuni  directly in faecal, feed, and environmental samples.    Results  With a sensitivity of 10 genome copies and a linear range of seven to eight orders of magnitude, the  C. coli  and  C. jejuni  real-time PCR assays allowed a precise quantification of purified DNA from  C. coli  and  C. jejuni . The assays were highly specific and showed a 6-log-linear dynamic range of quantification with a quantitative detection limit of approximately 2.5 × 10 2  CFU/g of faeces, 1.3 × 10 2  CFU/g of feed, and 1.0 × 10 3  CFU/m 2  for the environmental samples. Compared to the results obtained by culture, both  C. coli  and  C. jejuni  real-time PCR assays exhibited a specificity of 96.2% with a kappa of 0.94 and 0.89 respectively. For faecal samples of experimentally infected pigs, the coefficients of correlation between the  C. coli  or  C. jejuni  real-time PCR assay and culture enumeration were R 2  = 0.90 and R 2  = 0.93 respectively.    Conclusion  The  C. coli  and  C. jejuni  real-time quantitative PCR assays developed in this study provide a method capable of directly detecting and quantifying  C. coli  and  C. jejuni  in faeces, feed, and environmental samples. These assays represent a new diagnostic tool for studying the epidemiology of  Campylobacter  by, for instance, investigating the carriage and excretion of  C. coli  and  C. jejuni  by pigs from conventional herds.

Rapid identification and quantification of Campylobacter coli and Campylobacter jejuni by real-time PCR in pure cultures and in complex samples