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
2001
Cloak, O.M. | Duffy, G. | Sheridan, J.J. | Blair, I.S. | McDowell, D.A.
A rapid and sensitive method for the detection of Campylobacter in meat products was developed. Campylobacter were isolated from a range of Irish retail meats (n = 80) and poultry (n = 100) samples by direct plating on Campylobacter Selective Agar (CCDA, Oxoid). A total of 30-5% of samples tested positive for Campylobacter and Campylobacter jejuni was the most commonly identified species. The pathogen was detected in 39 (65%) poultry, 12 (60%) offal and 4 (20%) minced beef samples examined. Estimates derived from direct plate counts ranged from log10 0.7 to log10 2.75 cfu g(-1). The data from this study was used in the development of a rapid and sensitive method for 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 adhesion onto a polycarbonate membrane, phenol:chloroform extraction of DNA from the adherent 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 log10 4-5 and could be completed within 29 h. Results from SA-PCR analysis of a number of retail samples (n = 50) correlated favourably with traditional plate culture results, i.e. 34 samples were found to contain Campylobacter by both methods.
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