Overview
EN ISO 10272-2:2017/A1:2023 amends the horizontal ISO method for detection and enumeration of Campylobacter spp. by colony-count technique (Part 2). This amendment adds validated molecular confirmation and species identification options for thermotolerant Campylobacter (notably C. jejuni and C. coli, with reference to C. lari, C. upsaliensis and others) and revises performance testing requirements for culture media used in enumeration. The document is intended for food microbiology laboratories, regulatory bodies and organizations performing Campylobacter testing across the food chain.
Key topics and technical requirements
- Scope update: Inclusion of methods for molecular confirmation and identification of thermotolerant Campylobacter spp.; updated media performance testing.
- Colony-count fundamentals: Continued use of selective solid media (e.g., mCCD agar) and incubation in a microaerobic atmosphere at 41.5 °C for colony development.
- Biochemical vs molecular ID: Biochemical confirmation (catalase, hippurate, indoxyl acetate) remains described; Annex E provides molecular identification as an alternative.
- Multiplex real-time PCR (Annex D):
- Probe-based multiplex real-time PCR for confirmation of thermotolerant species (C. jejuni, C. coli, C. lari) targeting a 16S rRNA fragment.
- Includes reagent lists, oligonucleotide sequences (primers and probes), internal amplification control (IPC-ntb2 plasmid) and equipment requirements.
- Performance testing of culture media (Table B.1):
- Revised criteria and reference strains (WDCM catalogue) for productivity and selectivity of media like mCCD and Columbia blood agar.
- Example criterion: productivity ratio P ≥ 0.5 (see ISO 11133 for details on P).
- Definitions and notes: Updated definition of Campylobacter and emphasis on thermotolerant species relevant to human health.
- Validation and references: Method validation and inter-laboratory study results are noted (see Annex F and published Reference [12]).
Applications and users
- Food testing laboratories validating and performing Campylobacter enumeration and confirmation from food, primary production and environmental samples.
- Public health and regulatory agencies using standardized methods for surveillance, outbreak investigation and compliance.
- Diagnostic and contract labs integrating molecular confirmation (real-time PCR) to increase specificity and throughput.
- Producers and quality teams implementing HACCP and microbiological monitoring for poultry and other high‑risk commodities.
Related standards
Keywords: ISO 10272-2:2017/A1:2023, Campylobacter, colony-count technique, thermotolerant Campylobacter, multiplex real-time PCR, mCCD agar, culture media performance, food microbiology.