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BSI PD CEN ISO/TS 13136:2012

$142.49

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

Published By Publication Date Number of Pages
BSI 2012 32
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This Technical Specification describes the identification of Shiga toxin-producing Escherichia coli (STEC) by means of the detection of the following genes:

  1. the major virulence genes of STEC, stx and eae (References[2][3]);

  2. 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:

  1. products intended for human consumption and the feeding of animals;

  2. environmental samples in the area of food production and food handling;

  3. environmental samples in the area of primary production.

PDF Catalog

PDF Pages PDF Title
7 Introduction
8 1 Scope
2 Normative references
9 3 Terms and definitions
4 Principle
4.1 General
4.2 Microbial enrichment
10 4.3 Nucleic acid extraction
4.4 Target genes
4.5 Detection
4.6 Isolation
5 Diluents, culture media and reagents
11 5.1 Culture media
12 5.2 Reagents for nucleic acid extraction
5.3 Reagents for PCR
6 Equipment
13 7 Sampling
8 Preparation of test sample
9 Procedure
9.1 Test portion and initial suspension
14 9.2 Enrichment
9.3 Nucleic acid extraction
9.4 PCR amplification (for real-time PCR)
15 9.5 Strain isolation
10 Expression of results
11 Performance data
19 Annex A
(normative)

Flow diagram of the screening procedure

20 Annex B
(normative)

Flow diagram of the isolation and confirmation procedure

21 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

25 Annex D
(informative)

Internal amplification control

26 Annex E
(informative)

Primers and probes for the PCR assays

28 Annex F
(normative)

Isolation of STEC strains
Annex F
(normative)

Isolation of STEC strains

29 Bibliography
Bibliography
BSI PD CEN ISO/TS 13136:2012
$142.49