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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

Department of Veterinary Public Health, Faculty of Agriculture, Miyazaki University, 1-1 Gakuenkibanadai-nishi, Miyazaki 889-2192, Japan

Corresponding author: Dr N. Misawa (e-mail: a0d90lu{at}cc.miyazaki-u.ac.jp).

Received 9 May 2001; revised version received 15 July 2001; accepted 26 July 2001.

Abstract

A new combined filtration-enrichment culture followed by a PCR technique for the rapid detection of Campylobacter jejuni 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 10³5mucfu/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.

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