PCR-ELISAs for the detection of Campylobacter jejuni and Campylobacter coli in poultry samples

Campylobacter species, primarily Campylobacter jejuni and Campylobacter col i, are regarded as a major cause of human gastrointestinal disease, commonl y acquired by eating undercooked chicken. We describe a PCR-ELISA for the d etection of Campylobacter species and the discrimination of C. jejuni and C . coli in poultry samples. The PCR assay targets the 16S/23S ribosomal RNA intergenic spacer region of Campylobacter species with DNA oligonucleotide probes designed for the specific detection of C. jejuni, C. coli, and Campy lobacter species immobilized on NucleoLink(TM) ells and hybridized to PCR p roducts modified with a 5' biotin moiety. The limit of detection of the PCR -ELISA was 100-300 fg (40-120 bacterial cells) for C. jejuni and C. coli wi th their respective species-specific oligonucleotide probes and 10 fg (40-1 20 bacterial cells) with the Campylobacter genus-specific probe. Testing of poultry samples, which were presumptive positive for Campylobacter followi ng culture on the Malthus V analyzer with the PCR-ELISA determined Campylob acter to he present in 100% of samples (n = 40) with mixed cultures of C. j ejuni/C. coli in 55%. The PCR-ELISA when combined with culture pre-enrichme nt is able to detect the presence of Campylobacter and definitively identif y C. jejuni and C. coli in culture-enriched poultry meat samples.