Development of a combined filtration-enrichment culture followed by a one-step duplex PCR technique for the rapid detection of Campylobacter jejuni and C. coli in human faecal samples

A new combined filtration-enrichment culture followed by a PCR technique for the rapid detection of Campylobacterjejuni and C. coli in human faeces has been developed. Only bacteria that passed through the membrane could multiply in the enrichment culture; target bacteria were detected by a one-step duplex PCR technique with combinations of primers that are specific for different Campylobacter spp., which should allow for the detection of a mixed infection in a single patient. A Falcon cell-culture insert and 24-well tissue-culture plates were used. After 2 days, both C. jejuni and C. coli were reliably detected in diluted faeces that were seeded with as few as 10 cells which corresponds to a concentration of 10(3) cfu/g. Even allowing for the dilution of faecal samples, this represents an increase in sensitivity of two-to-three orders of magnitude over the conventional filtration method.