PD CEN ISO/TS 13136:2012
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
Hardcopy , PDF
English
30-11-2012
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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.
| Committee |
AW/9
|
| DocumentType |
Standard
|
| Pages |
32
|
| PublisherName |
British Standards Institution
|
| Status |
Current
|
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.
| 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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