XP CEN ISO/TS 13136

XP CEN ISO/TS 13136

December 2012
Standard Current

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

ISO/TR 13136:2012 describes 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.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.ISO/TR 13136:2012 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.

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National standards and national normative documents

Publication date

December 2012

Number of pages

35 p.

Reference

XP CEN ISO/TS 13136

ICS Codes

07.100.30   Food microbiology

Classification index

V08-790

Print number

1 - 26/12/2012

International kinship

ISO/TS 13136:2012

European kinship

CEN ISO/TS 13136:2012
Sumary
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

ISO/TR 13136:2012 describes 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.

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.

ISO/TR 13136:2012 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.

Table of contents
  • Avant-propos
    iv
  • Introduction
    v
  • 1 Domaine d'application
    1
  • 2 Références normatives
    2
  • 3 Termes et définitions
    2
  • 4 Principe
    2
  • 4.1 Généralités
    2
  • 4.2 Enrichissement microbien
    3
  • 4.3 Extraction des acides nucléiques
    3
  • 4.4 Gènes cibles
    3
  • 4.5 Détection
    3
  • 4.6 Isolement
    4
  • 5 Diluants, milieux de culture et réactifs
    4
  • 5.1 Milieux de culture
    4
  • 5.2 Réactifs pour l'extraction des acides nucléiques
    5
  • 5.3 Réactifs pour la PCR
    5
  • 6 Appareillage
    6
  • 7 Échantillonnage
    6
  • 8 Préparation de l'échantillon pour essai
    6
  • 9 Mode opératoire
    6
  • 9.1 Prise d'essai et suspension mère
    6
  • 9.2 Enrichissement
    7
  • 9.3 Extraction des acides nucléiques
    7
  • 9.4 Amplification par PCR (pour la PCR en temps réel)
    7
  • 9.5 Isolement de souches
    8
  • 10 Expression des résultats
    8
  • 11 Données de performances
    9
  • Annexe A (normative) Schéma du mode opératoire de criblage
    13
  • Annexe B (normative) Schéma du mode opératoire d'isolement et de confirmation
    14
  • Annexe C (informative) Identification des Escherichia coli producteurs de Shigatoxines (STEC) par amplification PCR multiplex des gènes de virulence et détection des amplicons par électrophorèse en gel d'agarose
    15
  • Annexe D (informative) Contrôle interne d'amplification
    20
  • Annexe E (informative) Amorces et sondes pour les réactions PCR
    21
  • Annexe F (normative) Isolement des souches STEC
    23
  • Bibliographie
    24
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