SIST EN 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 20837:2006)

This draft standard provides criteria and examples for sample preparation in order to obtain PCR-compatible samples or nucleic acids of suitable quality and quantity for PCR. The draft standard provides a description of the general principles involved followed by references to ISO-standards (Annex A) and detailed methods for DNA extraction (Annex B) and RNA extraction (Annex C). These methods can be applied to other food matrices, feed samples and agricultural/environmental matrices with some adaptations, if necessary.

Mikrobiologie von Lebensmitteln und Futtermitteln - Polymerase-Kettenreaktion (PCR) zum Nachweis von pathogenen Mikroorganismen in Lebensmitteln - Anforderungen an die Probenvorbereitung für den qualitativen Nachweis (ISO 20837:2006)

Diese Internationale Norm liefert Kriterien und Beispiele für die Probenvorbereitung, um für die PCR geeignete Proben oder Nukleinsäuren von für die PCR geeigneter Qualität und Quantität zu erhalten.
Sie liefert eine Beschreibung der in Frage kommenden allgemeinen Prinzipien, gefolgt von Verweisungen auf ISO-Normen (Anhang A) und einem ausführlich beschriebenen Verfahren der DNS-Extraktion (Anhang B).
Diese Internationale Norm wurde für Lebensmittelproben aufgestellt, gegebenenfalls kann sie auch mit einigen Anpassungen auf Futtermittel- und landwirtschaftliche/Umweltproben angewendet werden.

Microbiologie des aliments - Réaction de polymérisation en chaîne (PCR) pour la détection des micro-organismes pathogenes dans les aliments - Exigences relatives a la préparation des échantillons pour la détection qualitative (ISO 20837:2006)

L'ISO 20837:2006 fournit des critères et des exemples de préparation d'échantillons permettant d'obtenir des échantillons compatibles avec la méthode PCR ou des acides nucléiques de qualité appropriée et en quantité suffisante pour la méthode PCR.
Elle contient une description des principes généraux mis en oeuvre. Les références de normes relatives à l'enrichissement de micro-organismes sont données dans l'Annexe A et une méthode détaillée d'extraction de l'ADN est donnée dans l'Annexe B.
L'ISO 20837:2006 a été élaborée pour des matrices alimentaires mais pourrait également être appliquée à des aliments pour animaux et à des matrices agricoles/environnementales avec quelques adaptations, si nécessaire.

Mikrobiologija živil in krme – Odkrivanje prisotnosti patogenih mikroorganizmov v živilih z metodo verižne reakcije s polimerazo (PCR) – Zahteve za pripravo vzorca za kvalitativno odkrivanje (ISO 20837:2006)

General Information

• Status: Published
• Publication Date: 31-Aug-2006
• ICS: 07.100.30 - Food microbiology
• Technical Committee: KŽP - Agricultural food products
• Current Stage: 6060 - National Implementation/Publication (Adopted Project)
• Start Date: 01-Sep-2006
• Due Date: 01-Sep-2006
• Completion Date: 01-Sep-2006
• Ref Project: EN 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 20837:2006)

Standards Content (Sample)

SLOVENSKI STANDARD
SIST EN ISO 20837:2006
01-september-2006
Mikrobiologija živil in krme – Odkrivanje prisotnosti patogenih mikroorganizmov v
živilih z metodo verižne reakcije s polimerazo (PCR) – Zahteve za pripravo vzorca
za kvalitativno odkrivanje (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 20837:2006)
Mikrobiologie von Lebensmitteln und Futtermitteln - Polymerase-Kettenreaktion (PCR)
zum Nachweis von pathogenen Mikroorganismen in Lebensmitteln - Anforderungen an
die Probenvorbereitung für den qualitativen Nachweis (ISO 20837:2006)
Microbiologie des aliments - Réaction de polymérisation en chaîne (PCR) pour la
détection des micro-organismes pathogenes dans les aliments - Exigences relatives a la
préparation des échantillons pour la détection qualitative (ISO 20837:2006)
Ta slovenski standard je istoveten z: EN ISO 20837:2006
ICS:
07.100.30 Mikrobiologija živil Food microbiology
SIST EN ISO 20837:2006 en
2003-01.Slovenski inštitut za standardizacijo. Razmnoževanje celote ali delov tega standarda ni dovoljeno.

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EUROPEAN STANDARD
EN ISO 20837
NORME EUROPÉENNE
EUROPÄISCHE NORM
April 2006
ICS 07.100.30

English Version
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 20837:2006)
Microbiologie des aliments - Réaction de polymérisation en Mikrobiologie von Lebensmitteln und Futtermitteln -
chaîne (PCR) pour la détection des micro-organismes Polymerase-Kettenreaktion (PCR) zum Nachweis von
pathogènes dans les aliments - Exigences relatives à la pathogenen Mikroorganismen in Lebensmitteln -
préparation des échantillons pour la détection qualitative Anforderungen an die Probenvorbereitung bei qualitativem
(ISO 20837:2006) Nachweis (ISO 20837:2006)
This European Standard was approved by CEN on 13 April 2006.
CEN members are bound to comply with the CEN/CENELEC Internal Regulations which stipulate the conditions for giving this European
Standard the status of a national standard without any alteration. Up-to-date lists and bibliographical references concerning such national
standards may be obtained on application to the Central Secretariat or to any CEN member.
This European Standard exists in three official versions (English, French, German). A version in any other language made by translation
under the responsibility of a CEN member into its own language and notified to the Central Secretariat has the same status as the official
versions.
CEN members are the national standards bodies of Austria, Belgium, Cyprus, Czech Republic, Denmark, Estonia, Finland, France,
Germany, Greece, Hungary, Iceland, Ireland, Italy, Latvia, Lithuania, Luxembourg, Malta, Netherlands, Norway, Poland, Portugal, Romania,
Slovakia, Slovenia, Spain, Sweden, Switzerland and United Kingdom.
EUROPEAN COMMITTEE FOR STANDARDIZATION
COMITÉ EUROPÉEN DE NORMALISATION
EUROPÄISCHES KOMITEE FÜR NORMUNG
Management Centre: rue de Stassart, 36  B-1050 Brussels
© 2006 CEN All rights of exploitation in any form and by any means reserved Ref. No. EN ISO 20837:2006: E
worldwide for CEN national Members.

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EN ISO 20837:2006 (E)





Foreword


This document (EN ISO 20837:2006) has been prepared by Technical Committee CEN/TC 275
"Food analysis - Horizontal methods", the secretariat of which is held by DIN, in collaboration
with Technical Committee ISO/TC 34 "Agricultural food products".

This European Standard shall be given the status of a national standard, either by publication of
an identical text or by endorsement, at the latest by October 2006, and conflicting national
standards shall be withdrawn at the latest by October 2006.

According to the CEN/CENELEC Internal Regulations, the national standards organizations of
the following countries are bound to implement this European Standard: Austria, Belgium,
Cyprus, Czech Republic, Denmark, Estonia, Finland, France, Germany, Greece, Hungary,
Iceland, Ireland, Italy, Latvia, Lithuania, Luxembourg, Malta, Netherlands, Norway, Poland,
Portugal, Romania, Slovakia, Slovenia, Spain, Sweden, Switzerland and United Kingdom.


2

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INTERNATIONAL ISO
STANDARD 20837
First edition
2006-04-15

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
Microbiologie des aliments — Réaction de polymérisation en chaîne
(PCR) pour la détection des micro-organismes pathogènes dans les
aliments — Exigences relatives à la préparation des échantillons pour la
détection qualitative




Reference number
ISO 20837:2006(E)
©
ISO 2006

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ISO 20837:2006(E)
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ii © ISO 2006 – All rights reserved

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ISO 20837:2006(E)
Contents Page
Foreword. iv
Introduction . v
1 Scope. 1
2 Normative references. 1
3 Principle. 1
4 General laboratory requirements. 2
5 Reagents, apparatus and equipment. 2
6 Procedure. 2
Annex A (informative) Standards concerning the enrichment of microorganisms (bacteria) . 4
Annex B (informative) Method for DNA extraction from Gram-negative bacteria . 5
Bibliography . 7

© ISO 2006 – All rights reserved iii

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ISO 20837:2006(E)
Foreword
ISO (the International Organization for Standardization) is a worldwide federation of national standards bodies
(ISO member bodies). The work of preparing International Standards is normally carried out through ISO
technical committees. Each member body interested in a subject for which a technical committee has been
established has the right to be represented on that committee. International organizations, governmental and
non-governmental, in liaison with ISO, also take part in the work. ISO collaborates closely with the
International Electrotechnical Commission (IEC) on all matters of electrotechnical standardization.
International Standards are drafted in accordance with the rules given in the ISO/IEC Directives, Part 2.
The main task of technical committees is to prepare International Standards. Draft International Standards
adopted by the technical committees are circulated to the member bodies for voting. Publication as an
International Standard requires approval by at least 75 % of the member bodies casting a vote.
Attention is drawn to the possibility that some of the elements of this document may be the subject of patent
rights. ISO shall not be held responsible for identifying any or all such patent rights.
ISO 20837 was prepared by the European Committee for Standardization (CEN) Technical Committee
CEN/TC 275, Food analysis — Horizontal methods, in collaboration with Technical Committee ISO/TC 34,
Food products, Subcommittee SC 9, Microbiology, in accordance with the Agreement on technical
cooperation between ISO and CEN (Vienna Agreement).

iv © ISO 2006 – All rights reserved

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ISO 20837:2006(E)
Introduction
The detection of food-borne pathogens by PCR is usually performed by means of the following successive (or
simultaneous) steps:
⎯ homogenization of the sample;
⎯ (cultural) enrichment of the pathogen under study and sample treatment;
⎯ nucleic acid extraction (optional);
⎯ amplification of nucleic acids from the pathogen under study;
⎯ detection of the amplified DNA from the pathogen under study.
References to International Standards concerning enrichment of bacteria from food matrices are given in
Annex A. An example of a specific method for sample preparation is given in Annex B.
This International Standard is related to a series of standards and a Technical Specification under the general
title Microbiology of food and animal feeding stuffs — Polymerase chain reaction (PCR) for the detection of
food-borne pathogens:
⎯ General requirements and definitions (ISO 22174)
⎯ Requirements for sample preparation for qualitative detection (ISO 20837)
⎯ Performance testing for thermal cyclers (ISO/TS 20836)
⎯ Requirements for amplification and detection for qualitative methods (ISO 20838).
The International Organization for Standardization (ISO) draws attention to the fact that it is claimed that
compliance with this document may involve the use of one or more patents concerning the PCR technology.
ISO takes no position concerning the evidence, validity and scope of these patent rights.
ISO has been informed that Applied Biosystems, Roche Molecular Systems, Inc. and F. Hoffman-La Roche
Ltd. hold patent rights concerning the PCR technology. The companies have assured ISO that they are willing
to negotiate licences under reasonable and non-discriminatory terms and conditions with applicants
throughout the world. In this respect, the statements of the holders of these patent rights are registered with
ISO. Information may be obtained from:
Licensing Department
Applied Biosystems
850 Lincoln Centre Drive
Foster City, CA 94404
USA
and
Roche Molecular Systems, Inc.
Licensing Department
1145 Atlantic Avenue
Alameda, CA 94501
USA
© ISO 2006 – All rights reserved v

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ISO 20837:2006(E)
Attention is drawn to the possibility that some of the elements of this document may be the subject of patent
rights other than those identified above. ISO shall not be held responsible for identifying any or all such patent
rights.

vi © ISO 2006 – All rights reserved

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INTERNATIONAL STANDARD ISO 20837:2006(E)

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
WARNING — The use of this standard may involve hazardous materials, operations and equipment.
This standard does not purport to address all of the safety problems associated with its use. It is the
responsibility of the user of this standard to establish appropriate safety and health practices and
determine the applicability of regulatory limitations prior to use.
1 Scope
This International Standard provides criteria and examples for sample preparation in order to obtain PCR-
compatible samples or nucleic acids of suitable quality and quantity for PCR.
It provides a description of the general principles involved. References to standards concerning the
enrichment of microorganisms are given in Annex A, and a detailed method for DNA extraction is given in
Annex B.
This International Standard has been established for food matrices, but could also be applied to feed and
agricultural/environmental matrices with some adaptations, if necessary.
2 Normative references
The following referenced documents are indispensable for the application of this document. For dated
references, only the edition cited applies. For undated references, the latest edition of the referenced
document (including any amendments) applies.
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
3 Principle
3.1 General
The objective of the sample preparation methods described is to obtain samples or nucleic acids of suitable
quality and quantity for PCR.
NOTE The quality of nucleic acids depends for example on the chemical purity, the average length of the molecules
and the struct
...