A multiplex PCR and denaturing high-performance liquid chromatography method for identifying Campylobacter jejuni and Campylobacter coli

We report a new molecular technique for the analysis of Campylobacter jejuni and Campylobacter coli based on the separation of target fragments by denaturing high-performance liquid chromatography(DHPLC).A multiplex PCR(mPCR) assay for detection of Campylobacter jejuni and Campylobacter coli was developed using primers that specifically amplify segments of the 16S rRNA,gyrA and cdt genes.The mPCR had a detection limit of 10 pg/μL genomic DNA for Campylobacter jejuni and 1 pg/μL for Campylobacter coli.No cross-reaction was detected with non-thermo-tolerant Campylobacter and other 22 strains tested.The assay could detect 1.5 CFU/mL Campylobacter jejuni and Campylobacter coli in artificial inoculated chicken meat sample after enrichment at 42 ℃ in microaerophilic condition for 24 h.Test on 172 chicken meat samples detected 18 Campy-lobacter jejuni positive and 7 Campylobacter coli positive.These results indicated that the multiplex PCR-DHPLC assay could be used for specific detection of Campylobacter jejun and Campylobacter coli.