Development of a PCR ELISA assay for the identification of Campylobacter jejuni and Campylobacter coli

A polymerase chain reaction (PCR) assay was developed based on a solution-h ybridization colorimetric end-point detection format (PCR ELISA) for the id entification of Campylobacter jejuni and Campylobacter coli. PCR primers we re designed to target a gene sequence with species-specific motifs. Five bi otin-labelled probes targeted to the species-specific motifs were investiga ted for the detection of digoxygenin-labelled PCR products from C. jejuni a nd C coli using the PCR ELISA format. Two probes were identified, one which reacts with both the C. jejuni and C. coli target sequences (probe CC2) an d one probe which reacts with the C jejuni target sequence only (probe CJ2) . The specificity of the assay with the CJ2 and CC2 probes was investigated with a range of Campylobacter spp., Arcobacter spp., Helicobacter spp. and a range of unrelated organisms. The PCR ELISA assay and probes were demons trated to be specific for C. jejuni and C coli. The sensitivity of the PCR ELISA assay was demonstrated to be 10-100-fold more sensitive than a gel-ba sed PCR method using the same primers. This PCR ELISA assay is sensitive, s pecific and significantly reduces the time needed for the identification of C. jejuni and C. coli and has the potential to facilitate early detection of these important gastro-intestinal pathogens. (C) 2001 Academic Press.