CEN ISO/TS 18867:2015

Microbiology of the food chain - Polymerase chain reaction (PCR) for the detection of food-borne pathogens - Detection of pathogenic Yersinia enterocolitica and Yersinia pseudotuberculosis (ISO/DTS 18867:2015)

ISO/TS 18867:2015 specifies two horizontal methods for detection of the pathogenic bioserotypes of Y. enterocolitica and one for detection of Y. pseudotuberculosis by using real-time PCR-based methods. The described methods allow for the detection of the two pathogens in enrichments and allow the isolation of colonies. Y. pestis, the causative agent of bubonic and pneumonic plague harbours a variant of the ail gene as well and will be detected by the same primer/probe set as Y. pseudotuberculosis. However, Y. pestis is normally not associated with food. This Technical Specification is applicable to products for human consumption, animal feeding stuffs, and environmental samples.

Mikrobiologie der Lebensmittelkette - Polymerase-Kettenreaktion (PCR) zum Nachweis von pathogenen Mikroorganismen in Lebensmitteln - Nachweis von pathogenen Yersinia enterocolitica und Yersinia pseudotuberculosis (ISO/DTS 18867:2015)

Microbiologie de la chaîne alimentaire - Réaction de polymérisation en chaîne (PCR) pour la détection de micro-organismes pathogènes dans les aliments - Détection des Yersinia enterocolitica et Yersinia pseudotuberculosis pathogènes (ISO/DTS 18867:2015)

ISO/TS 18867:2015 décrit deux méthodes horizontales pour la détection de biosérotypes pathogènes des Y. enterocolitica ainsi qu'une méthode pour la détection des Y. pseudotuberculosis en faisant appel à des méthodes basées sur la PCR en temps réel. Les méthodes décrites permettent la détection des deux micro-organismes pathogènes par enrichissement et permettent d'isoler des colonies. Y. pestis, l'agent responsable de la peste bubonique et pneumonique héberge aussi un variant du gène ail et sera détecté par le même ensemble d'amorce et de sonde que Y. pseudotuberculosis. Toutefois, Y. pestis n'est normalement pas associé aux aliments. La présente Spécification technique est applicable aux produits destinés à la consommation humaine, aux aliments pour animaux et aux échantillons environnementaux.

Mikrobiologija v prehranski verigi - Polimerazna verižna reakcija (PCR) za ugotavljanje prisotnosti patogenih mikroorganizmov v živilih - Ugotavljanje prisotnosti patogenih Yersinia enterocolitica in Yersinia pseudotuberculosis (ISO/DTS 18867:2015)

Ta tehnična specifikacija določa dve horizontalni metodi za ugotavljanje prisotnosti patogenih mikroorganizmov Yersinia enterocolitica in ene metode za odkrivanje prisotnosti mikroorganizmov Yersinia pseudotuberculosis z uporabo polimerne verižne reakcije v realnem času. Metode ugotavljajo prisotnost dveh patogenov s polimerno verižno reakcijo in omogočajo osamitev kolonij. Y. pestis, povzročitelj bubonske in pljučne kuge, vsebuje tudi različico bolezenskega gena in ga zazna isti premaz/sonda, nastavljena kot Y. pseudotuberculosis. Vendar Y. pestis običajno ni povezan s hrano. Ta tehnična specifikacija se uporablja za proizvode za prehrano ljudi, živalsko krmo in okoljske vzorce.

General Information

• Status: Published
• Publication Date: 13-Oct-2015
• ICS: 07.100.30 - Food microbiology
• Technical Committee: CEN/TC 463 - Microbiology of the food chain
• Drafting Committee: CEN/TC 463/WG 1 - General requirements relating to PCR methods
• Current Stage: 6060 - Definitive text made available (DAV) - Publishing
• Start Date: 14-Oct-2015
• Due Date: 15-Mar-2015
• Completion Date: 14-Oct-2015
• Ref Project: SIST-TS CEN ISO/TS 18867:2015 - Microbiology of the food chain - Polymerase chain reaction (PCR) for the detection of food-borne pathogens - Detection of pathogenic Yersinia enterocolitica and Yersinia pseudotuberculosis (ISO/DTS 18867:2015)

Standards Content (Sample)

SLOVENSKI STANDARD
01-december-2015
Mikrobiologija v prehranski verigi - Polimerazna verižna reakcija (PCR) za
ugotavljanje prisotnosti patogenih mikroorganizmov v živilih - Ugotavljanje
prisotnosti patogenih Yersinia enterocolitica in Yersinia pseudotuberculosis
(ISO/DTS 18867:2015)
Microbiology of the food chain - Polymerase chain reaction (PCR) for the detection of
food-borne pathogens - Detection of pathogenic Yersinia enterocolitica and Yersinia
pseudotuberculosis (ISO/DTS 18867:2015)
Mikrobiologie der Lebensmittelkette - Polymerase-Kettenreaktion (PCR) zum Nachweis
von pathogenen Mikroorganismen in Lebensmitteln - Nachweis von pathogenen Yersinia
enterocolitica und Yersinia pseudotuberculosis (ISO/DTS 18867:2015)
Microbiologie de la chaîne alimentaire - Réaction de polymérisation en chaîne (PCR)
pour la détection de micro-organismes pathogènes dans les aliments - Détection des
Yersinia enterocolitica et Yersinia pseudotuberculosis pathogènes (ISO/DTS
18867:2015)
Ta slovenski standard je istoveten z: CEN ISO/TS 18867:2015
ICS:
07.100.30 Mikrobiologija živil Food microbiology
2003-01.Slovenski inštitut za standardizacijo. Razmnoževanje celote ali delov tega standarda ni dovoljeno.

CEN ISO/TS 18867
TECHNICAL SPECIFICATION
SPÉCIFICATION TECHNIQUE
October 2015
TECHNISCHE SPEZIFIKATION
ICS
English Version
Microbiology of the food chain - Polymerase chain reaction
(PCR) for the detection of food-borne pathogens -
Detection of pathogenic Yersinia enterocolitica and
Yersinia pseudotuberculosis (ISO/TS 18867:2015)
Microbiologie de la chaîne alimentaire - Réaction de Mikrobiologie der Lebensmittelkette - Polymerase-
polymérisation en chaîne (PCR) pour la détection de Kettenreaktion (PCR) zum Nachweis von pathogenen
micro-organismes pathogènes dans les aliments - Mikroorganismen in Lebensmitteln - Nachweis von
Détection des Yersinia enterocolitica et Yersinia pathogenen Yersinia enterocolitica und Yersinia
pseudotuberculosis pathogènes (ISO/TS 18867:2015) pseudotuberculosis (ISO/TS 18867:2015)
This Technical Specification (CEN/TS) was approved by CEN on 29 May 2015 for provisional application.

The period of validity of this CEN/TS is limited initially to three years. After two years the members of CEN will be requested to
submit their comments, particularly on the question whether the CEN/TS can be converted into a European Standard.

CEN members are required to announce the existence of this CEN/TS in the same way as for an EN and to make the CEN/TS
available promptly at national level in an appropriate form. It is permissible to keep conflicting national standards in force (in
parallel to the CEN/TS) until the final decision about the possible conversion of the CEN/TS into an EN is reached.

CEN members are the national standards bodies of Austria, Belgium, Bulgaria, Croatia, Cyprus, Czech Republic, Denmark, Estonia,
Finland, Former Yugoslav Republic of Macedonia, France, Germany, Greece, Hungary, Iceland, Ireland, Italy, Latvia, Lithuania,
Luxembourg, Malta, Netherlands, Norway, Poland, Portugal, Romania, Slovakia, Slovenia, Spain, Sweden, Switzerland, Turkey and
United Kingdom.
EUROPEAN COMMITTEE FOR STANDARDIZATION
COMITÉ EUROPÉEN DE NORMALISATION

EUROPÄISCHES KOMITEE FÜR NORMUNG

CEN-CENELEC Management Centre: Avenue Marnix 17, B-1000 Brussels
© 2015 CEN All rights of exploitation in any form and by any means reserved Ref. No. CEN ISO/TS 18867:2015 E
worldwide for CEN national Members.

Contents Page
European foreword . 3

European foreword
This document (CEN ISO/TS 18867:2015) has been prepared by Technical Committee ISO/TC 34 "Food
products" in collaboration with Technical Committee CEN/TC 275 “Food analysis - Horizontal methods”
the secretariat of which is held by DIN.
Attention is drawn to the possibility that some of the elements of this document may be the subject of
patent rights. CEN [and/or CENELEC] shall not be held responsible for identifying any or all such patent
rights.
According to the CEN-CENELEC Internal Regulations, the national standards organizations of the
following countries are bound to announce this Technical Specification: Austria, Belgium, Bulgaria,
Croatia, Cyprus, Czech Republic, Denmark, Estonia, Finland, Former Yugoslav Republic of Macedonia,
France, Germany, Greece, Hungary, Iceland, Ireland, Italy, Latvia, Lithuania, Luxembourg, Malta,
Netherlands, Norway, Poland, Portugal, Romania, Slovakia, Slovenia, Spain, Sweden, Switzerland,
Turkey and the United Kingdom.
Endorsement notice
The text of ISO/TS 18867:2015 has been approved by CEN as CEN ISO/TS 18867:2015 without any
modification.
TECHNICAL ISO/TS
SPECIFICATION 18867
First edition
2015-09-15
Microbiology of the food chain —
Polymerase chain reaction (PCR)
for the detection of food-borne
pathogens — Detection of pathogenic
Yersinia enterocolitica and Yersinia
pseudotuberculosis
Microbiologie de la chaîne alimentaire — Réaction de polymérisation
en chaîne (PCR) pour la détection de micro-organismes pathogènes
dans les aliments — Détection des Yersinia enterocolitica et Yersinia
pseudotuberculosis pathogènes
Reference number
ISO/TS 18867:2015(E)
©
ISO 2015
ISO/TS 18867:2015(E)
© ISO 2015, Published in Switzerland
All rights reserved. Unless otherwise specified, no part of this publication may be reproduced or utilized otherwise in any form
or by any means, electronic or mechanical, including photocopying, or posting on the internet or an intranet, without prior
written permission. Permission can be requested from either ISO at the address below or ISO’s member body in the country of
the requester.
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ii © ISO 2015 – All rights reserved

ISO/TS 18867:2015(E)
Contents Page
Foreword .v
Introduction .vi
1 Scope . 1
2 Normative references . 1
3 Terms and definitions . 1
4 Principles . 1
4.1 General . 1
4.2 Microbial enrichment . 2
4.3 Nucleic acid extraction . 2
4.4 Amplification and detection . 2
4.5 Isolation . 2
5 Reagents . 2
5.1 General . 2
5.2 Culture media . 2
5.2.1 General. 2
5.2.2 Diluent . 2
5.2.3 Enrichment media . 2
5.2.4 Selective solid medium . 4
5.2.5 Potassium hydroxide in saline solution, KOH . 5
5.3 Nucleic acid extraction . 5
5.4 Reagents for PCR . 5
5.5 Primers and probes. 5
6 Apparatus and equipment . 5
6.1 General . 5
6.2 Equipment for sample preparation prior to enrichment . 6
6.3 Equipment for microbial enrichment . . 6
6.4 Equipment for nucleic acid extraction . 6
6.5 Equipment for real-time PCR. 6
7 Sampling . 6
8 Procedure. 6
8.1 Sample preparation prior to enrichment . 6
8.1.1 General. 6
8.1.2 Preparation of the sample . 6
8.2 Microbial enrichment . 7
8.2.1 Pathogenic Y. enterocolitica . 7
8.2.2 Y. pseudotuberculosis . 7
8.2.3 Pathogenic Y. enterocolitica and Y. pseudotuberculosis . 7
8.3 Isolation of colonies, optional . 7
8.3.1 Pathogenic Y. enterocolitica . 7
8.3.2 Y. pseudotuberculosis . 7
8.3.3 Process controls . 8
8.4 Nucleic acid extraction .
...