1、November 2011 Translation by DIN-Sprachendienst.English price group 10No part of this translation may be reproduced without prior permission ofDIN Deutsches Institut fr Normung e. V., Berlin. Beuth Verlag GmbH, 10772 Berlin, Germany,has the exclusive right of sale for German Standards (DIN-Normen).I
2、CS 07.100.30!$vT(“1834905www.din.deDDIN EN ISO 22118Microbiology of food and animal feeding stuffs Polymerase chain reaction (PCR) for the detection and quantification offood-borne pathogens Performance characteristics (ISO 22118:2011)English translation of DIN EN ISO 22118:2011-11Mikrobiologie von
3、Lebensmitteln und Futtermitteln Polymerase-Kettenreaktion (PCR) zum Nachweis und zur quantitativen Bestimmung vonpathogenen Mikroorganismen in Lebensmitteln Leistungsmerkmale (ISO 22118:2011)Englische bersetzung von DIN EN ISO 22118:2011-11Microbiologie des aliments Raction de polymrisation en chane
4、 (PCR) pour la dtection et la quantification desmicro-organismes pathognes dans les aliments Caractristiques de performance (ISO 22118:2011)Traduction anglaise de DIN EN ISO 22118:2011-11www.beuth.deIn case of doubt, the German-language original shall be considered authoritative.Document comprises 1
5、6 pages11.11 DIN EN ISO 22118:2011-11 2 A comma is used as the decimal marker. National foreword This standard has been prepared by Technical Committee ISO/TC 34 “Food products” (Secretariat: AFNOR, France/ABNT, Brazil), Subcommittee SC 9 “Microbiology” (Secretariat: AFNOR, France), in collaboration
6、 with Technical Committee CEN/TC 275 “Food analysis Horizontal methods” (Secretariat: DIN, Germany), Working Group WG 6 “Microbiology of the food chain”. Based on the results of the parallel voting procedure carried out in accordance with the Agreement on technical cooperation between ISO and CEN (V
7、ienna Agreement), ISO 22118:2011 has been adopted as a European Standard. The responsible German body involved in its preparation was the Normenausschuss Lebensmittel und landwirtschaftliche Produkte (Food and Agricultural Products Standards Committee), Working Committee NA 057-01-06 AA Mikrobiologi
8、sche Lebensmitteluntersuchung einschlielich Schnellverfahren, NA 57-01-06-01 AK Polymerase Kettenreaktion zum Nachweis von Mikroorganismen. The DIN Standards corresponding to the International Standards referred to in this document are as follows: ISO 16140 DIN EN ISO 16140 ISO 22174 DIN EN ISO 2217
9、4 National Annex NA (informative) Bibliography DIN EN ISO 16140, Microbiology of food and animal feeding stuffs Protocol for the validation of alternative methods DIN EN ISO 22174, Microbiology of food and animal feeding stuffs Polymerase chain reaction (PCR) for the detection of food-borne pathogen
10、s General requirements and definitions EUROPEAN STANDARD NORME EUROPENNE EUROPISCHE NORM EN ISO 22118 July 2011 ICS 07.100.30 English Version Microbiology of food and animal feeding stuffs - Polymerase chain reaction (PCR) for the detection and quantification of food-borne pathogens - Performance ch
11、aracteristics (ISO 22118:2011) Microbiologie des aliments - Raction de polymrisation en chane (PCR) pour la dtection et la quantification des micro-organismes pathognes dans les aliments - Caractristiques de performance (ISO 22118:2011) Mikrobiologie von Lebensmitteln und Futtermitteln - Polymerase-
12、Kettenreaktion (PCR) zum Nachweis und zur quantitativen Bestimmung von pathogenen Mikroorganismen in Lebensmitteln - Leistungsmerkmale (ISO 22118:2011) This European Standard was approved by CEN on 14 July 2011. CEN members are bound to comply with the CEN/CENELEC Internal Regulations which stipulat
13、e 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 Stan
14、dard 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 nat
15、ional 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, Switze
16、rland and United Kingdom. EUROPEAN COMMITTEE FOR STANDARDIZATION COMIT EUROPEN DE NORMALISATION EUROPISCHES KOMITEE FR NORMUNG Management Centre: Avenue Marnix 17, B-1000 Brussels 2011 CEN All rights of exploitation in any form and by any means reserved worldwide for CEN national Members. Ref. No. E
17、N ISO 22118:2011: EEN ISO 22118:2011 (E) 2 Contents PageForeword3Introduction .41 Scope 52 Normative references 53 Terms and definitions .54 Performance characteristics of qualitative and quantitative detection methods .75 Performance characteristics for validation.96 Validation report 11Annex A (in
18、formative) Minimum list of microorganisms for use in the exclusivity tests for detection bacteria 13Bibliography 14DIN EN ISO 22118:2011-11 EN ISO 22118:2011 (E) 3 Foreword This document (EN ISO 22118:2011) has been prepared by Technical Committee CEN/TC 275 “Food analysis Horizontal methods”, the s
19、ecretariat 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 stan
20、dards 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 I
21、nternal 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, Lu
22、xembourg, Malta, Netherlands, Norway, Poland, Portugal, Romania, Slovakia, Slovenia, Spain, Sweden, Switzerland and the United Kingdom. DIN EN ISO 22118:2011-11 EN ISO 22118:2011 (E) 4 Introduction Molecular detection methods have been developed during the last few decades, and are now available for
23、 the majority of food-borne pathogens. Some of these methods have the potential for quantitative analysis. Although until now most methods have been based on the polymerase chain reaction (PCR) and real-time PCR, other molecular detection and quantification principles should be kept under considerat
24、ion. To compare molecular methods with conventional methods or with other principles, it is necessary to generate minimum requirements for performance characteristics of the methods to be developed. This International Standard is part of a series of documents under the general title Microbiology of
25、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, Microbio
26、logy 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 pat
27、hogens 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 fe
28、eding 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 definitio
29、ns 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
30、reaction (PCR)-based method DIN EN ISO 22118:2011-11 EN ISO 22118:2011 (E) 5 1 Scope This International Standard specifies minimum requirements of performance characteristics for the detection of nucleic acid sequences (DNA or RNA) by molecular methods. This International Standard applies to the det
31、ection of food-borne pathogens in foodstuffs and isolates obtained from them using molecular detection methods based on the polymerase chain reaction (PCR). This International Standard is also applicable, for example, to the detection of food-borne pathogens in environmental samples and in animal fe
32、eding 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 r
33、eferences, only the edition cited applies. For undated references, the latest edition of the referenced document (including any amendments) applies. ISO 16140:2003, Microbiology of food and animal feeding stuffs Protocol for the validation of alternative methods ISO 22174:2005, Microbiology of food
34、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 analyte component detected or measured by the method of a
35、nalysis NOTE 1 The analyte can be a microorganism or virus, its components or products. NOTE 2 Adapted from ISO 16140:2003, 3.4. 3.2 qualitative method method of analysis whose response is either the presence or absence of the analyte detected either directly or indirectly in a certain amount of sam
36、ple ISO 16140:2003, 3.5. DIN EN ISO 22118:2011-11 EN ISO 22118:2011 (E) 6 3.3 quantitative method method of analysis whose response is the amount of the analyte measured either directly or indirectly in a certain amount of sample NOTE Adapted from ISO 16140:2003, 3.6. 3.4 robustness testing food mic
37、robiology subjecting the proposed method to small procedural changes or environmental factors to determine what influence, if any, they have on method performance3.5 selectivity food microbiology measure of the inclusivity (detection of the target microorganism or virus) and exclusivity (non-detecti
38、on of non-target microorganisms or viruses) 3.6 sensitivity food microbiology measure of the lowest number of analyte cells, particles or molecules which can be detected in a single reaction 3.7 specificity capacity to exclusively recognize the target to be detected, distinguishing it from similar s
39、ubstances and impurities ISO 22174:2005, 3.6.4 3.8 trueness closeness of agreement between the expectation of a test result or a measurement result and a true value ISO 3534-2:20061, 3.3.3 3.9 detection limit limit of detection LOD lowest concentration or content of the target organism relative to t
40、he defined amount of matrix that can be consistently detected under the experimental conditions specified in the method NOTE Adapted from ISO 22174:2005, 3.1.8. 3.10 quantification limit limit of quantification LOQ food microbiology smallest amount of analyte (that is the lowest actual number of org
41、anisms), which can be measured and quantified with defined trueness and precision under the experimental conditions by the method under validation NOTE Adapted from ISO 16140:2003, 6.2.2.2.3. 3.11 precision closeness of agreement between independent test/measurement results obtained under stipulated
42、 conditions DIN EN ISO 22118:2011-11 EN ISO 22118:2011 (E) 7 NOTE 1 Precision depends only on the distribution of random errors and does not relate to the true value or the specified value. NOTE 2 The measure of precision is usually expressed in terms of imprecision and computed as a standard deviat
43、ion of the test results or measurement results. Less precision is reflected by a larger standard deviation. NOTE 3 Quantitative measures of precision depend critically on the stipulated conditions. Repeatability conditions and reproducibility conditions are particular sets of extreme stipulated cond
44、itions. ISO 3534-2:2006 1, 3.3.4 4 Performance characteristics of qualitative and quantitative detection methods 4.1 General A molecular detection method shall meet performance characteristics according to this International Standard. Information about the performance characteristics of the molecula
45、r detection method shall be made available. This includes specific information on the multi- or single-laboratory trial, including relevant information obtained during pre-validation of the method (e.g. variation of parameters, reagents). 4.2 Scope of the method The purpose of the method should be i
46、ndicated. Information regarding the intended use and the limitations of the methods shall be provided. In particular, the method provider shall indicate that the criteria set out in this International Standard are fulfilled. 4.3 Scientific basis An overview of the principles and references to releva
47、nt scientific publications should be provided. 4.4 Selectivity 4.4.1 Inclusivity test 4.4.1.1 General Empirical results from testing the method with the target microorganism or viruses should be provided. This testing should include all relevant variants or types of the microorganism or viruses acco
48、rding to the scope of the method (4.2). 4.4.1.2 Minimum requirements for specificity Sequence variants of the target microorganism or viruses should be detected with comparable amplification efficiency, even if sequence differences in the primer and/or probe binding sites are present as indicated by
49、 the scope of the method (4.2). If available, 50 strains of the specific target microorganism or virus should be tested. 4.4.2 Exclusivity test 4.4.2.1 General Empirical results from testing the method with non-target microorganisms or viruses should be provided. This testing should include both taxonomically closely related and not closely related microorganisms or viruses. The method should clearly distinguish between target and non-target microorganisms or viruses. DIN EN ISO 22118:2011-11 EN
