1、BRITISH STANDARD BS EN 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 The European Standard EN ISO 22174:2005 has the status of a British Standard ICS 07.100.30 BS EN ISO 221
2、74:2005 This British Standard was published under the authority of the Standards Policy and Strategy Committee on 24 March 2005 BSI 24 March 2005 ISBN 0 580 45673 0 National foreword This British Standard is the official English language version of EN ISO 22174:2005. It is identical with ISO 22174:2
3、004 The UK participation in its preparation was entrusted to Technical Committee AW/9, Microbiology, which has the responsibility to: A list of organizations represented on this committee can be obtained on request to its secretary. Cross-references The British Standards which implement internationa
4、l or European publications referred to in this document may be found in the BSI Catalogue under the section entitled “International Standards Correspondence Index”, or by using the “Search” facility of the BSI Electronic Catalogue or of British Standards Online. This publication does not purport to
5、include all the necessary provisions of a contract. Users are responsible for its correct application. Compliance with a British Standard does not of itself confer immunity from legal obligations. aid enquirers to understand the text; present to the responsible international/European committee any e
6、nquiries on the interpretation, or proposals for change, and keep the UK interests informed; monitor related international and European developments and promulgate them in the UK. Summary of pages This document comprises a front cover, an inside front cover, the EN ISO title page, the EN ISO forewor
7、d page, the ISO title page, pages ii to vi, pages 1 to 11 and a back cover. The BSI copyright notice displayed in this document indicates when the document was last issued. Amendments issued since publication Amd. No. Date Comments EUROPEAN STANDARD NORME EUROPENNE EUROPISCHE NORM EN ISO 22174 Febru
8、ary 2005 ICS 07.100.30 English version Microbiology of food and animal feeding stuffs - Polymerase chain reaction (PCR) for the detection of food-borne pathogens - General requirements and definitions (ISO 22174:2005) Microbiologie des aliments - Raction de polymrisation en chane (PCR) pour la reche
9、rche de micro-organismes pathognes dans les aliments - Exigences gnrales et dfinitions (ISO 22174:2005) Mikrobiologie von Lebensmitteln und Futtermitteln - Polymerase- Kettenreaktion (PCR) zum Nachweis von pathogenen Mikroorganismen in Lebensmitteln - Allgemeine Anforderungen und Begriffe (ISO 22174
10、:2005) This European Standard was approved by CEN on 20 October 2004. 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 bibliographi
11、cal 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
12、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,
13、Lithuania, Luxembourg, Malta, Netherlands, Norway, Poland, Portugal, Slovakia, Slovenia, Spain, Sweden, Switzerland and United Kingdom. EUROPEAN COMMITTEE FOR STANDARDIZATION COMIT EUROPEN DE NORMALISATION EUROPISCHES KOMITEE FR NORMUNG Management Centre: rue de Stassart, 36 B-1050 Brussels 2005 CEN
14、 All rights of exploitation in any form and by any means reserved worldwide for CEN national Members. Ref. No. EN ISO 22174:2005: ENE ISO 122002:475 E() 2 Foreword This document (EN ISO 22174:2005) has been prepared by Technical Committee CEN/TC 275 “Food analysis - Horizontal methods“, the secretar
15、iat 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 August 2005, and conflicting national
16、 standards shall be withdrawn at the latest by August 2005. 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, G
17、ermany, Greece, Hungary, Iceland, Ireland, Italy, Latvia, Lithuania, Luxembourg, Malta, Netherlands, Norway, Poland, Portugal, Slovakia, Slovenia, Spain, Sweden, Switzerland and United Kingdom. ENISO22174:2005 Reference number ISO 22174:2005(E) OSI 5002INTERNATIONAL STANDARD ISO 22174 First edition
18、2005-02-15 Microbiology of food and animal feeding stuffs Polymerase chain reaction (PCR) for the detection of food-borne pathogens General requirements and definitions Microbiologie des aliments Raction de polymrisation en chane (PCR) pour la recherche de micro-organismes pathognes dans les aliment
19、s Exigences gnrales et dfinitions ENISO22174:2005IS:47122 O5002(E) DPlcsid Fremia ihTs PDF file may ctnoian emdebt dedyfepcaes. In ccaocnadrw eith Aebods licensilop gnic,y this file mairp eb ynted iv roweb detu slahl ton ide ebtlnu deess the typefaces whice era hml era deddebicsnede to i dnanstlaled
20、 t noeh computfrep reormign tide ehtin.g In wodlnidaot gnhis file, trapise atpecc tiereht nser ehnopsiiblity fo not infriigngn Aebods licensilop gnic.y ehT ISO tneClar Secrteiraat caceptl on siibality in this .aera Ai ebods a tredamafo kr Aebod SystemI sncotaropr.de teDails fo teh softwacudorp erts
21、sut deo crtaee this PDF file cna f ebi dnuon tlareneG eh Info leratit evo the file; tP ehDc-Frtaeion marapterew setpo erimizde for irpnti.gn Evyre caer neeb sah taken to sneeru that the file is suitlbae fosu re yb ISO memdob rebeis. In tlnu ehikletneve y ttah lborp aem leratit gno it is f,dnuo plsae
22、e inform ttneC ehlar Secrteiraat ta the serddaig sleb nevwo. ISO 5002 All irthgs erse.devr lnUeto sswrehise specified, on trap fo this lbupictaion maeb y cudorperro de tuilizi den yna form ro na ybm ynae,s lecetrinoc ro mecinahcal, inclidung tohpcoiypodna gn micrfoilm, wittuoh repmissii non writign
23、from ietI rehSa Ot tsserdda eh ebolw or ISOs member body in the country of ttseuqer ehe.r ISO cirypothg fofice saCe tsopale 65 eneG 1121-HC 02 av leT. 4 + 10 947 22 1 11 xaF0 947 22 14 + 9 74 E-mail coirypthgiso.o gr We bwww.is.o gro Pulbisdehi n Switlrez dnaii ISO 5002 Allr ithgsr esedevrENISO22174
24、:2005IS:47122 O5002(E) I SO 5002 All irthgs ersedevr iiiContents Page Foreword iv Introduction v 1 Scope 1 2 Normative references. 1 3 Terms and definitions. 1 4 Principle. 6 4.1 General. 6 4.2 Preliminary microbial enrichment . 6 4.3 Nucleic acid preparation 6 4.4 PCR amplification. 6 4.5 Detection
25、 and confirmation of PCR products 7 5 Test material 7 6 General laboratory requirements 7 6.1 General. 7 6.2 Personnel. 7 6.3 Laboratory setup. 7 6.4 Waste management 8 7 Reagents 8 8 Apparatus and equipment 8 8.1 General. 8 8.2 Special considerations. 8 9 Procedure. 8 9.1 Sample preparation. 8 9.2
26、Amplification. 9 9.3 Control reaction 9 9.4 Confirmation of PCR results 9 10 Evaluation 10 11 Test report 10 Bibliography . 11 ENISO22174:2005IS:47122 O5002(E) iv I SO 5002 All irthgs ersedevrForeword ISO (the International Organization for Standardization) is a worldwide federation of national stan
27、dards 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 org
28、anizations, 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
29、 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
30、 % of the member bodies casting a vote. ISO 22174 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
31、 the Agreement on technical cooperation between ISO and CEN (Vienna Agreement). ENISO22174:2005IS:47122 O5002(E) I SO 5002 All irthgs ersedevr vIntroduction The polymerase chain reaction (PCR) is a fast, sensitive and specific method for the detection of food-borne pathogens. Although a relatively y
32、oung technology, the application of PCR-based methods in food analysis is increasing. In brief, existing protocols can be divided in two main groups, depending on the type of nucleic acid used as target for amplification: RNA-based amplification (RT-PCR); DNA-based amplification (PCR). Numerous vari
33、ations of both methods have been established and can be characterized by their degree of complexity and automation. The level of specificity of the methods varies from screening assays which detect nucleic acid sequences common to a microbiological genus, to specific assays which identify nucleic ac
34、id sequences unique to an individual strain- or type-specific nucleic acid sequence. This International Standard presents a comprehensive list of requirements for PCR-based methods used for the detection of microorganisms in food samples. It contains terms and definitions used in reference to PCR an
35、d RT-PCR. ISO 22174 is part of a series of International 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); Requir
36、ements for sample preparation for qualitative detection (ISO 20837)1) ; Requirements for amplification and detection for qualitative methods (ISO 20838)1) ; Performance testing for thermal cyclers (ISO/TS 20836)1) . The International Organization for Standardization (ISO) draws attention to the fact
37、 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
38、F. Hoffman-La Roche Ltd. hold patent rights concerning the PCR technology. The companies have assured the 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
39、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 1) To be published. ENISO22174:2005IS:47122 O5002(E) vi I SO 5002 All irthgs ersedevrand Roche Molecular Systems, Inc. Licensin
40、g Department 1145 Atlantic Avenue Alameda, CA 94501 USA 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. ENISO22174:
41、2005INTENRATIONAL TSANDADR IS:47122 O5002(E)I SO 5002 All irthgs ersedevr 1Microbiology of food and animal feeding stuffs Polymerase chain reaction (PCR) for the detection of food-borne pathogens General requirements and definitions WARNING The use of this standard may involve hazardous materials, o
42、perations 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 determines the applicability of regulatory limitations prior to use. 1
43、 Scope This International Standard gives the general requirements for the in vitro amplification of nucleic acid sequences (DNA or RNA). It is applicable to the testing of foodstuffs and isolates obtained from foodstuffs for food-borne pathogens using the polymerase chain reaction (PCR). The minimum
44、 requirements laid down in this International Standard are intended to ensure that comparable and reproducible results are obtained in different laboratories. This International Standard has been established for food-borne pathogens in or isolated from food and feed matrices, but is also applicable
45、to other matrices (e.g. environmental samples) and for the detection of non- pathogenic microorganisms. 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
46、latest edition of the referenced document (including any amendments) applies. ISO 3534-1, Statistics Vocabulary and symbols Part 1: Probability and general statistical terms ISO 5725-1, Accuracy (trueness and precision) of measurement methods and results Part 1: General principles and definitions IS
47、O 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
48、of food-borne pathogens Requirements for amplification and detection for qualitative methods ISO/IEC 17025, General requirements for the competence of testing and calibration laboratories 3 Terms and definitions For the purposes of this document, the following terms and definitions apply. For defini
49、tions concerning validation, see ISO 3534-1 and ISO 5725-1. ENISO22174:2005ISO 22174:2005(E) 2 I SO 5002 All irthgs ersedevr3.1 General terms 3.1.1 nucleic acid macromolecule that is the medium for genetic information or acts as an agent in expressing the information NOTE There are two types of nucleic acid, DNA and RNA. 3.1.2 DNA deoxyribonucleic acid polymer of deoxyribonucleotides occurring in a doub
