Evaluation of a PCR assay for the detection and identification of Campylobacter jejuni and Campylobacter coli in retail poultry products

Abstract

A PCR-based method was applied to Campylobacter detection in poultry samples at the retail level. In total, 73 retail poultry samples purchased from supermarkets in the Basque Country area in the north of Spain were examined using both culture and molecular (alternative) methods. In our routine method, the worldwide ISO 10272:1995 standard of Preston broth incubated at 42 °C for conventional Campylobacter detection was adopted. The molecular method was comprised of a DNA extraction kit consisting of a single polypropylene spin column and PCR amplification of the Campylobacter 16S rRNA gene. A total of 54 raw samples were positive by either PCR or culture; among these, 50 were found to be positive by conventional plating and 54 by PCR. Concordant results, i.e., positive and negative in both methods, were found in 64 samples (94.1%). All positive samples by culture were also positive by PCR, resulting in 100% of positive concordance. Two samples (2.9%) positive after retesting by PCR were considered to be false-negatives. The detection limit of the PCR method was 5 CFUs that corresponded to 0.2 CFUs per 5 μl in the PCR mixture. The percentages of samples that required enrichment to prove Campylobacter presence were moderate, 18% by culture and 13% by PCR. Total analysis time was reduced to a few hours (within the working day) or 24 h when enrichment was required. Therefore, this PCR method proved to be useful as a routine diagnostic test for Campylobacter detection and confirmation of C. jejuni and C. coli in naturally contaminated poultry samples.