Validation of PCR methods for confirmation and species identification of thermotolerant Campylobacter as part of EN ISO 10272 - Microbiology of the food chain - Horizontal method for detection and enumeration of Campylobacter spp.

This article describes the outline and organization of the validation of three multiplex PCR methods for species identification and/or confirmation of thermotolerant Campylobacter spp. The three PCR methods were validated against the reference method described in the EN ISO standard 10272:2017 – Microbiology of the food chain – Horizontal method for detection and enumeration of Campylobacter. The results of the PCR methods were compared against the reference method in a method comparison study and an interlaboratory study, in large outlined according to EN ISO 16140-6:2019 – Microbiology of the food chain – Method validation Part 6: Protocol for the validation of alternative (proprietary) methods for microbiological confirmation and typing procedures. The performance, in terms of inclusivity and exclusivity, of each of the eight PCR targets were comparable to that of the reference method: close, equal, or better depending on the target. In total, all three PCR methods were concluded to be equally qualified as the reference method for molecular identification and/or confirmation of thermotolerant Campylobacter spp. C. jejuni, C. coli and C. lari isolated from the food chain and have been included in Amendment 1 of ISO 10272:2017.