PCR identification of Campylobacter coli and Campylobacter jejuni by partial sequencing of virulence genes

The objective of this study was to utilize a multiplex PCR assay for concurrent detection of Campylobacter spp. and C. coli or C. jejuni, using probes derived from genes cadF and ceuE and an undefined virulence gene. A total of 97 Campylobacter strains, isolated from turkey litter (n=74), chicken livers (n=15) and clinical (n=8) samples, were speciated using the PCR-based assay. PCR amplification of the isolates identified a 400-bp cadF gene, conserved in Campylobacter species, an 894-bp ceuE gene, specific for C. coli, and a 160-bp oxidoreductase gene, specific for C. jejuni. The ∼35 kDa cadF adhesion proteins allow Campylobacter to bind to the intestinal epithelial cells and the 37 kDa ceuE lipoproteins are involved in siderophore transport. Sequencing of the 160-bp undefined gene yielded a 67% protein identical match with a gene encoding an oxidoreductase subunit in C. jejuni. The specificity of the assay was validated on 36 non-Campylobacter strains (11 Gram-positive and 25 Gram-negative bacteria). The PCR assay identified 59% of turkey and 47% of chicken isolates as C. jejuni, and 41% of turkey and 53% of chicken isolates as C. coli. All human isolates were identified as C. jejuni. The specificity of this assay to detect C. coli or C. jejuni was 97%.