prEN ISO 13136-1
(Main)Microbiology of the food chain - Detection, isolation and characterization of Shiga toxin-producing Escherichia coli (STEC) - Part 1: Horizontal method for the detection and isolation of Shiga toxin-producing Escherichia coli (STEC) (ISO/DIS 13136-1:2024)
Microbiology of the food chain - Detection, isolation and characterization of Shiga toxin-producing Escherichia coli (STEC) - Part 1: Horizontal method for the detection and isolation of Shiga toxin-producing Escherichia coli (STEC) (ISO/DIS 13136-1:2024)
This standard describes the detection and isolation of Shiga toxin-producing Escherichia coli (STEC). The procedure includes the detection by Real Time PCR of the major virulence genes of STEC, stx1 and stx2 (Reference [2]) in enrichment cultures. Isolation of STEC from the enrichment culture is attempted if one or both stx genes are detected in the screening.
This document is applicable to
- products intended for human consumption,
- products intended for animal feeding,
- environmental samples in the area of food and feed production, handling, and
- samples from the primary production stage.
Mikrobiologie der Lebensmittelkette - Nachweis, Isolierung und Charakterisierung von Shiga-Toxin bildenden Escherichia coli (STEC) - Teil 1: Horizontales Verfahren zum Nachweis und zur Isolierung von Shiga-Toxin bildenden Escherichia coli (STEC) (ISO/DIS 13136‑1:2024)
Microbiologie de la chaîne alimentaire - Détection, isolement et caractérisation des Escherichia coli producteurs de Shigatoxines (STEC) - Partie 1: Méthode horizontale pour la détection et l’isolement des Escherichia coli producteurs de Shigatoxines (STEC (ISO/DIS 13136-1:2024)
Mikrobiologija v prehranski verigi - Odkrivanje prisotnosti, izolacija in karakterizacija Escherichia coli (STEC), ki proizvaja Shiga toksin - 1. del: Horizontalna metoda za odkrivanje prisotnosti in izolacijo Escherichia coli (STEC), ki proizvaja Shiga toksin (ISO/DIS 13136-1:2024)
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Standards Content (Sample)
SLOVENSKI STANDARD
01-april-2024
Mikrobiologija v prehranski verigi - Odkrivanje prisotnosti, izolacija in
karakterizacija Escherichia coli (STEC), ki proizvaja Shiga toksin - 1. del:
Horizontalna metoda za odkrivanje prisotnosti in izolacijo Escherichia coli (STEC),
ki proizvaja Shiga toksin (ISO/DIS 13136-1:2024)
Microbiology of the food chain - Detection, isolation and characterization of Shiga toxin-
producing Escherichia coli (STEC) - Part 1: Horizontal method for the detection and
isolation of Shiga toxin-producing Escherichia coli (STEC) (ISO/DIS 13136-1:2024)
Mikrobiologie der Lebensmittelkette - Nachweis, Isolierung und Charakterisierung von
Shiga-Toxin bildenden Escherichia coli (STEC) - Teil 1: Horizontales Verfahren zum
Nachweis und zur Isolierung von Shiga-Toxin bildenden Escherichia coli (STEC)
(ISO/DIS 13136-1:2024)
Microbiologie de la chaîne alimentaire - Détection, isolement et caractérisation des
Escherichia coli producteurs de Shigatoxines (STEC) - Partie 1: Méthode horizontale
pour la détection et l’isolement des Escherichia coli producteurs de Shigatoxines (STEC
(ISO/DIS 13136-1:2024)
Ta slovenski standard je istoveten z: prEN ISO 13136-1
ICS:
07.100.30 Mikrobiologija živil Food microbiology
2003-01.Slovenski inštitut za standardizacijo. Razmnoževanje celote ali delov tega standarda ni dovoljeno.
DRAFT
International
Standard
ISO/DIS 13136-1
ISO/TC 34/SC 9
Microbiology of the food
Secretariat: AFNOR
chain — Detection, isolation and
Voting begins on:
characterization of Shiga toxin-
2024-02-14
producing Escherichia coli (STEC) —
Voting terminates on:
2024-05-08
Part 1:
Horizontal method for the detection
and isolation of Shiga toxin-
producing Escherichia coli (STEC)
ICS: 07.100.30
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Reference number
ISO/DIS 13136-1:2024(en)
DRAFT
ISO/DIS 13136-1:2024(en)
International
Standard
ISO/DIS 13136-1
ISO/TC 34/SC 9
Microbiology of the food
Secretariat: AFNOR
chain — Detection, isolation and
Voting begins on:
characterization of Shiga toxin-
producing Escherichia coli (STEC) —
Voting terminates on:
Part 1:
Horizontal method for the detection
and isolation of Shiga toxin-
producing Escherichia coli (STEC)
ICS: 07.100.30
THIS DOCUMENT IS A DRAFT CIRCULATED
FOR COMMENTS AND APPROVAL. IT
IS THEREFORE SUBJECT TO CHANGE
AND MAY NOT BE REFERRED TO AS AN
INTERNATIONAL STANDARD UNTIL
PUBLISHED AS SUCH.
This document is circulated as received from the committee secretariat.
IN ADDITION TO THEIR EVALUATION AS
BEING ACCEPTABLE FOR INDUSTRIAL,
© ISO 2024
TECHNOLOGICAL, COMMERCIAL AND
USER PURPOSES, DRAFT INTERNATIONAL
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Published in Switzerland Reference number
ISO/DIS 13136-1:2023(en)
ii
ISO/DIS 13136-1:2023(en)
Contents Page
Foreword .iv
Introduction .v
1 Scope . 1
2 Normative references . 1
3 Terms and definitions . 2
4 Principle . 2
4.1 General .2
4.2 Microbial enrichment .2
4.3 Nucleic acid extraction .2
4.4 Target genes .3
4.5 Detection and isolation .3
5 Culture media and reagents . 3
6 Equipment and consumables . 3
7 Sampling . 4
8 Preparation of test sample . 5
9 Procedure . 5
9.1 Test portion and initial suspension .5
9.2 Enrichment .5
9.3 Nucleic acid extraction .6
9.4 PCR amplification (for real-time PCR) .6
9.4.1 General .6
9.4.2 Interpretation of real-time PCR results .6
9.4.3 Internal Amplification control for real-time PCR .6
9.5 STEC strain isolation .7
10 Expression of the results . 8
11 Validation of the method . 9
12 Test report . 9
13 Quality assurance . 10
Annex A (normative) Flow diagram of the procedure .11
Annex B (normative) Culture media and reagents .12
Annex C (normative) Flow diagram of the isolation of STEC strains .16
Annex D (normative) Primers and probes for the real-time PCR assays . 17
Annex E (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 .18
Annex F (informative) Acid treatment of enrichment cultures .22
Annex G (informative) Detection of Escherichia coli producing the Stx2f subtype by real-time
[5]
PCR (extract of the EURL-VTEC_Method_10 ) .23
Bibliography .25
iii
ISO/DIS 13136-1:2023(en)
Foreword
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This document was prepared by Technical Committee ISO/TC 34, Food products, Subcommittee SC 9,
Microbiology, in collaboration with the European Committee for Standardization (CEN) Technical Committee
CEN/TC 463, Microbiology of the food chain, in accordance with the
...
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