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PD CEN ISO/TS 13136:2012

Current
Current

The latest, up-to-date edition.

Microbiology of food and animal feed. Real-time polymerase chain reaction (PCR)-based method for the detection of food-borne pathogens. Horizontal method for the detection of Shiga toxin-producing Escherichia coli (STEC) and the determination of O157, O111, O26, O103 and O145 serogroups
Available format(s)

Hardcopy , PDF

Language(s)

English

Published date

30-11-2012

Foreword
Introduction
1 Scope
2 Normative references
3 Terms and definitions
4 Principle
5 Diluents, culture media and reagents
6 Equipment
7 Sampling
8 Preparation of test sample
9 Procedure
10 Expression of results
11 Performance data
Annex A (normative) - Flow diagram of the
        screening procedure
Annex B (normative) - Flow diagram of the
        isolation and confirmation procedure
Annex C (informative) - Identification of Shiga
        toxin-producing Escherichia coli
        (STEC) by multiplex PCR amplification
        of virulence genes and detection of PCR
        products by agarose gel electrophoresis
Annex D (informative) - Internal amplification control
Annex E (informative) - Primers and probes for the
        PCR assays
Annex F (normative) - Isolation of STEC strains
Bibliography

Specifies the identification of Shiga toxin-producing Escherichia coli (STEC) by means of the detection of the following genes: a) the major virulence genes of STEC, stx and eae; b) the genes associated with the serogroups O157, O111, O26, O103, and O145.

This Technical Specification describes the identification of Shiga toxin-producing Escherichia coli (STEC) by means of the detection of the following genes: the major virulence genes of STEC, stx and eae (References[2][3]); the genes associated with the serogroups O157, O111, O26, O103, and O145 (References[3][4]). In any case, when one or both of the stx genes is/are detected, the isolation of the strain is attempted. The isolation of STEC from samples positive for the presence of the genes specifying the serogroups in the scope of this method can be facilitated by using serogroup-specific enrichment techniques (e.g. immunomagnetic separation, IMS). The protocol uses real-time PCR as the reference technology for detection of the virulence and serogroup-associated genes. This Technical Specification is applicable to: products intended for human consumption and the feeding of animals; environmental samples in the area of food production and food handling; environmental samples in the area of primary production.

Committee
AW/9
DocumentType
Standard
Pages
32
PublisherName
British Standards Institution
Status
Current

Standards Relationship
CEN ISO/TS 13136:2012 Identical
ISO/TS 13136:2012 Identical

ISO 22174:2005 Microbiology of food and animal feeding stuffs Polymerase chain reaction (PCR) for the detection of food-borne pathogens General requirements and definitions
ISO 16140-2:2016 Microbiology of the food chain Method validation Part 2: Protocol for the validation of alternative (proprietary) methods against a reference method
ISO/TS 19036:2006 Microbiology of food and animal feeding stuffs Guidelines for the estimation of measurement uncertainty for quantitative determinations
ISO 7218:2007 Microbiology of food and animal feeding stuffs General requirements and guidance for microbiological examinations
ISO 7550:1985 Laboratory glassware — Disposable micropipettes
ISO 20837:2006 Microbiology of food and animal feeding stuffs Polymerase chain reaction (PCR) for the detection of food-borne pathogens Requirements for sample preparation for qualitative detection
ISO 20838:2006 Microbiology of food and animal feeding stuffs Polymerase chain reaction (PCR) for the detection of food-borne pathogens Requirements for amplification and detection for qualitative methods
ISO 16140:2003 Microbiology of food and animal feeding stuffs Protocol for the validation of alternative methods
ISO 16654:2001 Microbiology of food and animal feeding stuffs — Horizontal method for the detection of Escherichia coli O157

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