A survey on the incidence of Campylobacter spp. and the development of a surface adhesion polymerase chain reaction (SA-PCR) assay for the detection of Campylobacter jejuni in retail meat products

A rapid and sensitive method for the detection of Campylobacter in meat pro ducts was developed. Campylobacter were isolated from a range of Irish reta il meats (n = 80) and poultry (n = 100) samples by direct plating on Campyl obacter Selective Agar (CCDA, Oxoid). A total of 30.5% of samples tested po sitive for Campylobacter and Campylobacter jejuni was the most commonly ide ntified species. The pathogen was defected in 39 (65%) poultry, 12 (60%) of fal and 4 (20%) minced beef samples examined: Estimates derived from direct plate counts ranged from log(10) 0.7 to log(10) 2.75 cfu g(-1) The data fr om this study was used in the development of a rapid and sensitive method f or the detection of Campylobacter in poultry. A rapid method was developed based on an initial sample enrichment for 24 h in Campylobacter Enrichment Broth (CEB) recovery of the pathogen from the enriched sample by surface ad hesion onto a polycarbonate membrane, phenol:chloroform extraction of DNA f rom she ad herent bacteria, and PCR analysis using primers specific for the flagellin A gene (present in C, jejuni and C. coli). The developed surface adhesion PCR (SA-PCR) technique had a detection limit of log(10) 4-5 and c ould be completed within 29h. Results from SA-PCR analysis of a number of r etail samples (n=50) correlated favourably with traditional plate culture r esults, i.e. 34 samples were found to contain Campylobacter by both methods . (C) 2001 Academic Press.