Optimisation of specific PCR detection of Campylobacter coli in enrichment broth

Campylobacter jejuni and C. coli are among the most important causes of acute diarrhoea in humans throughout the world. Poultry meat is a major source of Campylobacter infections. Sensitive detection methods are necessary to identify contaminated samples. Detection of campylobacters by culturing is slow and tedious, whereas PCR technology offers the potential for rapid and sensitive detection, however, it may be inhibited when used directly for food or pre-enriched food samples. Different methods for sample and/or DNA preparation were studied to find an optimal combination for sensitive PCR detection of C. coli in enrichment broth. Buoyant density centrifugation (BDC) prior to cell lysis improved PCR detection of C. coli by 100-1000-fold. Preston enrichment broth spiked with 101-102 CFU ml-1 was detected positive after 18 h of enrichment. Specific flaA PCR detection of C. coli in enrichment broth following BDC and simple heat lysis of the cells can be conducted within two working days. This study is a par...