ISO 10272-2:2017/Amd 1:2023 PDF

Overview

ISO 10272-2:2017/Amd 1:2023 updates the international horizontal method for detection and enumeration of Campylobacter spp. using the colony‑count technique. This amendment integrates molecular confirmation and identification methods for thermotolerant Campylobacter (notably C. jejuni, C. coli, C. lari) and revises performance testing requirements for selective culture media. The document is intended for laboratories performing microbiological analysis of the food chain, public‑health surveillance and food safety testing.

Key topics

• Scope of amendment: Addition of molecular confirmation and species identification methods and changes to culture‑media performance testing.
• Colony‑count technique basics: Continued use of selective solid media (e.g., modified Charcoal Cefoperazone Deoxycholate (mCCD) agar), incubation in a microaerobic atmosphere at 41.5 °C, and biochemical/microscopy confirmation.
• Selective enrichment: Notes that thermotolerant Campylobacter typically grow in Bolton broth and Preston broth.
• Biochemical confirmation: Describes traditional tests (catalase, hippurate hydrolysis, indoxyl acetate) and updated differentiation (Table 2) among C. jejuni, C. coli and other species.
• Molecular methods (Annexes D & E):
  • Annex D: Multiplex real‑time PCR (probe‑based) targeting 16S rRNA for confirmation of thermotolerant Campylobacter spp.; includes reagent lists, primers/probe sequences, PCR setup, controls and a recommended temperature–time programme. Validation data showed true positives produced Cq values below 38.
  • Annex E: Additional molecular identification approaches as alternatives to biochemical identification.
  • Annex F: Interlaboratory validation summary for the PCR method.
• Performance testing of media (Table B.1): Updated criteria for productivity and selectivity using reference strains (WDCM catalogue) and references to method validation standards.

Applications and users

• Food microbiology laboratories performing routine testing and enumeration of Campylobacter in poultry, dairy, produce and other food matrices.
• Public health and clinical labs involved in outbreak investigation and species confirmation.
• Food industry QC and contract testing labs using standardized methods for regulatory compliance and supplier verification.
• Method developers and validation laboratories applying performance testing and interlaboratory study requirements.

Related standards

• ISO 22174 (nucleic acid methods - reagent quality and PCR guidance)
• ISO 11133 (microbiological culture media - performance criteria)
• ISO 16140‑6 (method validation, interlaboratory studies)
• ISO 6887 series (sample preparation for microbiological examination)

This amendment strengthens ISO 10272-2 by combining traditional colony counts with validated molecular confirmation, improving specificity and harmonizing culture‑media performance - essential for reliable Campylobacter testing across the food chain.