SIST EN ISO 22119:2011
(Main)Microbiology of food and animal feeding stuffs - Real-time polymerase chain reaction (PCR) for the detection of food-borne pathogens - General requirements and definitions (ISO 22119:2011)
Microbiology of food and animal feeding stuffs - Real-time polymerase chain reaction (PCR) for the detection of food-borne pathogens - General requirements and definitions (ISO 22119:2011)
This standard is applicable to the detection of food-borne pathogens in foodstuffs, and isolates obtained from them, using the polymerase chain reaction (PCR). The standard describes requirements for the amplification and detection of nucleic acid sequences (DNA or RNA after reverse transcription) by real-time PCR. The minimum requirements laid down in the standard provide the basis for comparable
and reproducible results in individual and between different laboratories.
NOTE The standard can also be used for other purposes in the foodstuffs sector (e.g. the detection of food-borne pathogens in environmental samples), and for animal feeding stuffs.
NOTE Because of the rapid progress in this field, the examples given are those most frequently used at time when the standard was developed.
Mikrobiologie von Lebensmitteln und Futtermitteln - Real-Time-Polymerase-Kettenreaktion (PCR) für den Nachweis von pathogenen Mikroorganismen in Lebensmitteln - Allgemeine Anforderungen und Begriffe (ISO 22119:2011)
Die vorliegende Norm gilt für den Nachweis von pathogenen Mikroorganismen in Lebensmitteln und in aus Lebensmitteln isolierten Bakterienisolaten unter Anwendung der Polymerase-Kettenreaktion (PCR). Die Norm beschreibt die Anforderungen an die Amplifikation (Vervielfältigung) und den Nachweis von Nukleinsäure¬sequenzen (DNS oder RNS nach Umschreibung durch reverse Transkription) durch Real-Time-PCR.
Die in dieser Norm festgelegten Mindestanforderungen liefern die Grundlage für vergleichbare und reprodu¬zierbare Ergebnisse in einzelnen Laboratorien oder zwischen verschiedenen Laboratorien.
ANMERKUNG 1 Die Norm kann auch für andere Zwecke im Lebensmittelsektor (z. B. zum Nachweis von über Nahrungsmittel übertragenen pathogenen Mikroorganismen in Umgebungsproben oder für Tierfuttermittel angewendet werden.
ANMERKUNG 2 Auf Grund des schnellen Fortschritts auf diesem Gebiet entsprechen die angegebenen Beispiele den zum Zeitpunkt der Erarbeitung dieser Norm häufigsten Anwendungen.
Microbiologie des aliments - Réaction de polymérisation en chaîne (PCR) en temps réel pour la détection des micro-organismes pathogènes dans les aliments - Exigences générales et définitions (ISO 22119:2011)
L'ISO 22119:2010 définit les termes relatifs à la détection de micro-organismes pathogènes présents dans les aliments et les extraits obtenus à partir de ces aliments, en faisant appel à la réaction de polymérisation en chaîne (PCR). L'ISO 22119:2010 spécifie également les exigences relatives à l'amplification et à la détection des séquences d'acide nucléique (ADN ou ARN après transcription inverse) par une PCR en temps réel.
Les exigences minimales spécifiées dans l'ISO 22119:2010 servent de base pour l'obtention de résultats comparables et reproductibles au sein d'un laboratoire et dans des laboratoires différents.
L'ISO 22119:2010 peut également être utilisée, par exemple, pour la détection de micro-organismes pathogènes d'origine alimentaire dans des prélèvements environnementaux ainsi que dans les aliments pour animaux.
Mikrobiologija živil in krme - Odkrivanje prisotnosti patogenih mikroorganizmov v živilih s polimerazno verižno reakcijo (PCR) v realnem času - Splošne zahteve in definicije (ISO 22119:2011)
Ta mednarodni standard določa izraze za odkrivanje prisotnosti patogenih mikroorganizmov v živilih in izolatih, pridobljenih iz njih, z uporabo polimerazne verižne reakcije (PRC). Ta mednarodni standard tudi opredeljuje zahteve za povečevanje in odkrivanje prisotnosti zaporedij nukleinskih kislin (DNK ali RNK po reverzni transkripciji) s PRC v realnem času. Minimalne zahteve, določene v tem mednarodnem standardu, zagotavljajo temelj za primerljive in ponovljive rezultate v posameznem laboratoriju in med različnimi laboratoriji. Ta mednarodni standard se uporablja tudi, na primer, za odkrivanje prisotnosti patogenih mikroorganizmov v živilih v okoljskih vzorcih in v krmah.
General Information
Standards Content (Sample)
SLOVENSKI STANDARD
SIST EN ISO 22119:2011
01-oktober-2011
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åLYLOLKVSROLPHUD]QRYHULåQRUHDNFLMR 3&5 YUHDOQHPþDVX6SORãQH]DKWHYHLQ
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Microbiology of food and animal feeding stuffs - Real-time polymerase chain reaction
(PCR) for the detection of food-borne pathogens - General requirements and definitions
(ISO 22119:2011)
Mikrobiologie von Lebensmitteln und Futtermitteln - Real-Time-Polymerase-
Kettenreaktion (PCR) für den Nachweis von pathogenen Mikroorganismen in
Lebensmitteln - Allgemeine Anforderungen und Begriffe (ISO 22119:2011)
Microbiologie des aliments - Réaction de polymérisation en chaîne (PCR) en temps réel
pour la détection des micro-organismes pathogènes dans les aliments - Exigences
générales et définitions (ISO 22119:2011)
Ta slovenski standard je istoveten z: EN ISO 22119:2011
ICS:
07.100.30 Mikrobiologija živil Food microbiology
SIST EN ISO 22119:2011 en
2003-01.Slovenski inštitut za standardizacijo. Razmnoževanje celote ali delov tega standarda ni dovoljeno.
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SIST EN ISO 22119:2011
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SIST EN ISO 22119:2011
EUROPEAN STANDARD
EN ISO 22119
NORME EUROPÉENNE
EUROPÄISCHE NORM
July 2011
ICS 07.100.30
English Version
Microbiology of food and animal feeding stuffs - Real-time
polymerase chain reaction (PCR) for the detection of food-borne
pathogens - General requirements and definitions (ISO
22119:2011)
Microbiologie des aliments - Réaction de polymérisation en Mikrobiologie von Lebensmitteln und Futtermitteln - Real-
chaîne (PCR) en temps réel pour la détection des micro-
time-Polymerase-Kettenreaktion (PCR) zum Nachweis von
organismes pathogènes dans les aliments - Exigences pathogenen Mikroorganismen in Lebensmitteln -
générales et définitions (ISO 22119:2011) Allgemeine Anforderungen und Begriffe (ISO 22119:2011)
This European Standard was approved by CEN on 14 July 2011.
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 CEN-CENELEC Management Centre 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 CEN-CENELEC Management Centre has the same
status as the official versions.
CEN members are the national standards bodies of Austria, Belgium, Bulgaria, Croatia, 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: Avenue Marnix 17, B-1000 Brussels
© 2011 CEN All rights of exploitation in any form and by any means reserved Ref. No. EN ISO 22119:2011: E
worldwide for CEN national Members.
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SIST EN ISO 22119:2011
EN ISO 22119:2011 (E)
Contents Page
Foreword .3
2
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SIST EN ISO 22119:2011
EN ISO 22119:2011 (E)
Foreword
This document (EN ISO 22119:2011) 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 "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 January 2012, and conflicting national standards shall be withdrawn at
the latest by January 2012.
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 implement this European Standard: Austria, Belgium, Bulgaria, Croatia, 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 the United Kingdom.
3
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SIST EN ISO 22119:2011
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SIST EN ISO 22119:2011
INTERNATIONAL ISO
STANDARD 22119
First edition
2011-07-15
Microbiology of food and animal feeding
stuffs — Real-time polymerase chain
reaction (PCR) for the detection of food-
borne pathogens — General
requirements and definitions
Microbiologie des aliments — Réaction de polymérisation en chaîne
(PCR) en temps réel pour la détection des micro-organismes
pathogènes dans les aliments — Exigences générales et définitions
Reference number
ISO 22119:2011(E)
©
ISO 2011
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SIST EN ISO 22119:2011
ISO 22119:2011(E)
COPYRIGHT PROTECTED DOCUMENT
© ISO 2011
All rights reserved. Unless otherwise specified, no part of this publication may be reproduced or utilized in any form or by any means,
electronic or mechanical, including photocopying and microfilm, without permission in writing from either ISO at the address below or
ISO's member body in the country of the requester.
ISO copyright office
Case postale 56 • CH-1211 Geneva 20
Tel. + 41 22 749 01 11
Fax + 41 22 749 09 47
E-mail copyright@iso.org
Web www.iso.org
Published in Switzerland
ii © ISO 2011 – All rights reserved
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SIST EN ISO 22119:2011
ISO 22119:2011(E)
Contents
Foreword .iv
Introduction.v
1 Scope.1
2 Normative references.1
3 Terms and definitions .1
4 Principle.5
4.1 General .5
4.2 Probes for real-time PCR.5
5 General laboratory requirements.6
6 Reagents and materials .6
7 Apparatus.7
8 Laboratory sample .8
9 Procedure.8
9.1 Sample preparation .8
9.2 Amplification.8
9.3 Controls.9
9.4 Analysis of the fluorescence data .9
10 Evaluation and documentation .11
Bibliography.12
© ISO 2011 – All rights reserved iii
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SIST EN ISO 22119:2011
ISO 22119:2011(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 22119 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 2011 – All rights reserved
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SIST EN ISO 22119:2011
ISO 22119:2011(E)
Introduction
The polymerase chain reaction (PCR) has been shown to be a fast, sensitive, and specific method for
detection of food-borne pathogens. Further developments of the technology allow the detection of specific
PCR products generated by the amplification process. The principle relies on the excitation of fluorescent
markers during the PCR process.
This International Standard is part of a series of documents under the general title Microbiology of food and
animal feeding stuffs — Polymerase chain reaction (PCR) for the detection of food-borne pathogens:
ISO/TS 20836, Microbiology of food and animal feeding stuffs — Polymerase chain reaction (PCR) for the
detection of food-borne pathogens — Performance testing for thermal cyclers
ISO 20837, 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 20838, Microbiology of food and animal feeding stuffs — Polymerase chain reaction (PCR) for the
detection of food-borne pathogens — Requirements for amplification and detection for qualitative methods
ISO 22118, Microbiology of food and animal feeding stuffs — Polymerase chain reaction (PCR) for the
detection and quantification of food-borne pathogens — Performance characteristics
ISO 22119, Microbiology of food and animal feeding stuffs — Real-time polymerase chain reaction (PCR) for
the detection of food-borne pathogens — General requirements and definitions
ISO 22174, Microbiology of food and animal feeding stuffs — Polymerase chain reaction (PCR) for the
detection of food-borne pathogens — General requirements and definitions
The following Technical Specification is in preparation:
ISO/TS 13136, Microbiology of food and animal feeding stuffs — Horizontal method for the detection of Shiga
toxin-producing Escherichia coli (STEC) belonging to O157, O111, O26, O103 and O145 serogroups —
Qualitative real-time polymerase chain reaction (PCR)-based method
© ISO 2011 – All rights reserved v
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SIST EN ISO 22119:2011
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SIST EN ISO 22119:2011
INTERNATIONAL STANDARD ISO 22119:2011(E)
Microbiology of food and animal feeding stuffs — Real-time
polymerase chain reaction (PCR) for the detection of food-
borne pathogens — General requirements and definitions
1 Scope
This International Standard defines terms for the detection of food-borne pathogens in foodstuffs, and isolates
obtained from them, using the polymerase chain reaction (PCR). This International Standard also specifies
requirements for the amplification and detection of nucleic acid sequences (DNA or RNA after reverse
transcription) by real-time PCR.
The minimum requirements laid down in this International Standard provide the basis for comparable and
reproducible results within individual and between different laboratories.
This International Standard is also applicable, for example, to the detection of food-borne pathogens in
environmental samples and in animal feeding stuffs.
NOTE Because of the rapid progress in this field, the examples given are those most frequently in use at the time of
development of this International Standard.
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 20838, Microbiology of food and animal feeding stuffs — Polymerase chain reaction (PCR) for the
detection of food-borne pathogens — Requirements for amplification and detection for qualitative methods
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 Terms and definitions
For the purposes of this document, the following terms and definitions apply.
3.1
real-time polymerase chain reaction
real-time PCR
enzymatic procedure which combines the in vitro amplification of specific DNA segments by a process of
denaturation, annealing of specific primers, and synthesis of DNA with the detection of specific PCR products
during the amplification process
NOTE 1 Generally, the amplification reaction mixture contains one or more specific DNA probes coupled with one or
more fluorescent dyes. Using this technology, the signal is generated after specific hybridization of the probes to the target
nucleic acid sequence and excitation with light of a definite wavelength.
© ISO 2011 – All rights reserved 1
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SIST EN ISO 22119:2011
ISO 22119:2011(E)
NOTE 2 The use of non-specific DNA-binding fluorescent dyes can be applied if positive results are verified in
accordance with ISO 20838.
3.2
PCR product
DNA amplified by PCR
[ISO 22174:2005, 3.4.5]
3.3
fluorescence resonance energy transfer
FRET
〈food-borne pathogen detection by PCR〉 distance-dependent energy transfer from a donor molecule to an
acceptor molecule resulting in enhanced fluorescence of the acceptor molecule after excitation with
electromagnetic radiation of a definite wavelength
NOTE Taken from Reference [2].
3.4
reporter
〈food-borne pathogen detection by PCR〉 fluorescent molecule used to detect the hybridization of specific
probes by excitation with electromagnetic radiation of an appropriate wavelength
3.5
quencher
〈food-borne pathogen detection by PCR〉 fluorescent molecule serving as an energy acceptor and thus
quenching the fluorescence signal of the reporter (donor)
3.6
dark quencher
molecule serving as an acceptor, which does not emit energy in a spectral range detected by the optical
detection system of the real-time PCR instrument
3.7
5′-3′-exonuclease activity
ability of an enzyme, e.g. a nucleic acid polymerase, to cleave a hybridized nucleic acid molecule in the 5′-3′-
direction
NOTE The activity of 5′-3′-exonuclease is double stranded DNA specific. It is dependent on the type of enzyme and
can be present, for example, in Taq-, Tth- and Tfl-polymerase.
3.8
fluorescent probe
oligonucleotide or oligonucleotide analogon of defined sequence coupled with one or more fluorescent
molecules
NOTE Any system emitting a fluorescence signal after specific hybridization to the target nucleic acid sequence
which can be detected by the specific equipment can be used as a fluorescent probe.
3.9
hydrolysis probe
fluorescent probe coupled with two fluorescent molecules which are sterically separated by the 5′-3′-
exonuclease activity of the enzyme during the amplification process
NOTE The pr
...
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