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A PCR assay for the detection of Campylobacter jejuni and Campylobacter coli in water
Author(s) -
Kirk R.,
Rowe M.T.
Publication year - 1994
Publication title -
letters in applied microbiology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.698
H-Index - 110
eISSN - 1472-765X
pISSN - 0266-8254
DOI - 10.1111/j.1472-765x.1994.tb00460.x
Subject(s) - campylobacter jejuni , campylobacter , sonication , microbiology and biotechnology , campylobacter coli , lysis , membrane filter , biology , chromatography , polymerase chain reaction , bacteria , chemistry , membrane , gene , biochemistry , genetics
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 fla 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 spiked 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 .