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.
Kirk
R
R
Department of Agriculture for Northern Ireland, Belfast, UK.
Rowe
M T
MT
D017000:Campylobacter coli / Q000302:isolation & purification*; D016123:Campylobacter jejuni / Q000302:isolation & purification*; D005374:Filtration; D008855:Microscopy, Electron, Scanning; D016133:Polymerase Chain Reaction / Q000379:methods*; D012680:Sensitivity and Specificity; D013010:Sonication; D014871:Water Microbiology