ISO INTERNATIONAL STANDARD 22174 First edition 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 Réaction de polymérisation en chaine (PCR) pour la recherche de micro-organismes pathogenes dans les aliments Exigences genérales et definitions Reference number ISO 22174:2005(E) @ ISO 2005 Copyright International Organization for Standardization ted without license from IHS Not for Resale ISO 22174:2005(E) PDF disclaimer This PDF file may contain embedded typefaces. In accordance with Adobe's licensing policy, this file may be printed or viewed but shall not be edited unless the typefaces which are embedded are licensed to and installed on the computer performing the editing. In accepts no liability in this area. Adobe is a trademarkof AdobeSystems Incorporated. Details of the software products used to create this PDF file can be found in the General Info relative to the file; the PDF-creation parameters were optimized for printing. Every care has been taken to ensure that the file is suitable for use by IsO member bodies. In the unlikely event that a problem relating to it is found, please inform the Central Secretariat at the address given below ?ISO2005 All rights reserved. Unless otherwise specified, no part of this publication may be reproduced or utilized in any form or by any means, 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 [email protected] Web www.iso.org Published in Switzerland yemainagazainandardizato @ISO2005-All rightsreserved mittedwithoutlicense from IHS Not for Resale ISO 22174:2005(E) Contents Page Foreword Introduction 1 Scope 2 Normative references 3 Terms and definitions 4 Principle 4.1 General... 4.2 Preliminary microbial enrichment . 4.3 Nucleic acid preparation ..... 4.4 PCR amplification ... 4.5 Detection and confirmation of PcR products... 5 Test material 6 General laboratory requirements . 6.1 General.... 6.2 Personnel.... 6.3 Laboratory setup.. 6.4 Waste management. 7 Reagents. 8 Apparatus and equipment. 8.1 8 8.2 Special considerations 9 Procedure.... 8 9.1 Sample preparation 9.2 Amplification. 6 9.3 Control reaction .... 9.4 Confirmation of PCR results 10 Evaluation 10 11 Test repor 10 Bibliography Copyrght International Organizaionforstandardizaon'ghts reserved ili Not for Resale ISO 22174:2005(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 technicalcommittees.Eachmemberbodyinterestedinasubjectforwhichatechnicalcommitteehasbeen InternationalElectrotechnicalCommission(iEC)onallmattersofelectrotechnicalstandardization International Standards are drafted in accordance with the rules given in the ISO/lEC Directives, Part 2. 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. 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 the Agreement on technical cooperation between ISO and CEN (Vienna Agreement). ional OrganizationforStandardizatior @ISO2005-All rightsreserved ISO 22174:2005(E) Introduction The polymerase chain reaction (PCR) is a fast, sensitive and specific method for the detection of food-borne pathogens. Although a relatively young 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 nucle