A PCR assay for the detection of Campylobacter jejuni and Campylobacter coli in water

A PCR assay has been developed for the detection of Campylobacter jejuni and Camp. coli in water samples. The sample is filtered through a membrane which is subjected to sonication to release the impacted cells. After removal of the filter from the cell suspension and a freeze/thaw cell lysis step, a semi-nested PCR is carried out on the filtrate using the primers CF02, CF03 and CF04 (Camp. jejuni flaA and flaB gene sequences). Incorporation of a sonication stage allows removal of the filter membrane since they have been shown to inhibit the PCR. In experiments with spike water samples (20 ml) a theoretical sensitivity of 10-20 campylobacter cells ml-1 was achieved. Using a sample volume of 100 ml this sensitivity can be increased to approximately 2 campylobacter cells ml-1.