SIST-TS CEN ISO/TS 13136:2013 PDF

Overview

CEN ISO/TS 13136:2012 (ISO/TS 13136:2012) is a technical specification for the microbiology of food and animal feed that defines a real-time PCR (qPCR)–based, horizontal method for the detection of Shiga toxin-producing Escherichia coli (STEC). The method targets the principal STEC virulence genes (stx) and the locus of enterocyte effacement gene (eae), and it provides procedures to determine the presence of the serogroups O157, O111, O26, O103 and O145.

Key topics

• Target genes and markers
  • Detection of stx (Shiga toxin) and eae (attachment factor) genes as the primary virulence markers.
  • Detection of serogroup-associated genetic markers for O157, O111, O26, O103, O145.
• Reference technology
  • Uses real-time PCR as the reference detection technology for virulence and serogroup-associated genes.
• Enrichment and isolation
  • Describes sample enrichment, nucleic acid extraction and qPCR amplification workflows.
  • Specifies that when one or both stx genes are detected, an attempt to isolate the strain must be made.
  • Recommends serogroup-specific enrichment/isolation aids (e.g., immunomagnetic separation, IMS) to facilitate recovery of STEC strains.
• Sample scope
  • Applicable to foods for human consumption and animal feed, and to environmental samples in food production, handling and primary production.
• Supporting material
  • Includes flow diagrams for screening and isolation, annexes on isolation procedures, internal amplification controls, and lists of primers/probes (informative/ normative annexes).

Applications

• Food safety testing and surveillance
  • Screening of ready-to-eat foods, raw ingredients and animal feed for STEC contamination using a standardized qPCR workflow.
• Environmental monitoring
  • Testing of food-processing environments, water and primary-production samples for STEC presence.
• Outbreak investigation and public health labs
  • Rapid molecular screening to prioritize samples for culture-based confirmation and strain characterization.
• Quality assurance in industry
  • Integrating an ISO/TS-based detection method into HACCP, supplier verification and microbiological control plans.

Who should use it

• Food microbiology laboratories, public health and veterinary diagnostic labs, regulatory agencies, food producers and contract testing labs seeking a validated qPCR-based screening method for STEC serogroups of high public‑health importance.

Related standards

• This document complements other ISO and CEN standards on food microbiology methods, enrichment media and culture-based confirmation. Users should consult applicable national/regional regulatory standards when implementing testing programs.

Keywords: CEN ISO/TS 13136:2012, ISO/TS 13136, STEC detection, real-time PCR, Shiga toxin-producing Escherichia coli, O157 O111 O26 O103 O145, food microbiology, immunomagnetic separation, isolation, stx eae.