1、November 2016 English price group 27No 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).ICS 07.100.30!%uj“2588271www.din.de
2、DIN EN ISO 16140-2Microbiology of the food chain Method validation Part 2: Protocol for the validation of alternative (proprietary) methods against a reference method (ISO 161402:2016);English version EN ISO 161402:2016,English translation of DIN EN ISO 16140-2:2016-11Mikrobiologie der Lebensmittelk
3、ette Verfahrensvalidierung Teil 2: Arbeitsvorschrift fr die Validierung von alternativen (urheberrechtlich geschtzten) Verfahren anhand eines Referenzverfahrens (ISO 161402:2016);Englische Fassung EN ISO 161402:2016,Englische bersetzung von DIN EN ISO 16140-2:2016-11Microbiologie de la chane aliment
4、aire Validation des mthodes Partie 2: Protocole pour la validation de mthodes alternatives (commerciales) par rapport une mthode de rfrence (ISO 161402:2016);Version anglaise EN ISO 161402:2016,Traduction anglaise de DIN EN ISO 16140-2:2016-11Together with DIN EN ISO 161401:201611,supersedesDIN EN I
5、SO 16140:201112www.beuth.deDocument comprises 75 pagesDTranslation by DIN-Sprachendienst.In case of doubt, the German-language original shall be considered authoritative.11.16 DIN EN ISO 16140-2:2016-11 2 A comma is used as the decimal marker. National foreword This document (EN ISO 16140-2:2016) ha
6、s been prepared by Technical Committee ISO/TC 34 “Food products”, Subcommittee SC 9 “Microbiology” in collaboration with Technical Committee CEN/TC 275 “Food analysis Horizontal methods”, Working Group WG 6 “Microbiology of the food chain” (Secretariat: AFNOR, France). The responsible German body in
7、volved in its preparation was DIN-Normenausschuss Lebensmittel und landwirtschaftliche Produkte (DIN Standards Committee Food and Agricultural Products), Working Committee NA 057-01-12 AA Validierung mikrobiologischer Verfahren. The DIN Standards corresponding to the International Standards referred
8、 to in this document are as follows: ISO 5725-2 DIN ISO 5725-2 ISO 11133 DIN EN ISO 11133 ISO 16140-1 DIN EN ISO 16140-1 ISO 16297 DIN EN ISO 16297 ISO/TS 22117 DIN CEN ISO/TS 22117, DIN SPEC 91215 Amendments This standard differs from DIN EN ISO 16140:2011-12 as follows: a) the standard has been re
9、vised and divided into several parts; b) the present part deals with the protocol for the validation of alternative methods against a reference method; c) the standard has been editorially revised. Previous editions DIN EN ISO 16140: 2003-09, 2011-12 DIN EN ISO 16140-2:2016-11 3 National Annex NA (i
10、nformative) Bibliography DIN ISO 5725-2, Accuracy (trueness and precision) of measurement methods and results Part 2: Basic method for the determination of repeatability and reproducibility of a standard measurement method DIN EN ISO 11133, Microbiology of food, animal feed and water Preparation, pr
11、oduction, storage and performance testing of culture media DIN EN ISO 16140-1, Microbiology of the food chain Method validation Part 1: Vocabulary DIN EN ISO 16297, Milk Bacterial count Protocol for the evaluation of alternative methods DIN CEN ISO/TS 22117, DIN SPEC 91215, Microbiology of food and
12、animal feeding stuffs Specific requirements and guidance for proficiency testing by interlaboratory comparison DIN EN ISO 16140-2:2016-11 4 This page is intentionally blank EUROPEAN STANDARD NORME EUROPENNE EUROPISCHE NORM EN ISO 16140-2 July 2016 ICS 07.100.30 Supersedes EN ISO 16140:2003English Ve
13、rsion Microbiology of the food chain - Method validation - Part 2: Protocol for the validation of alternative (proprietary) methods against a reference method (ISO 16140-2:2016)Microbiologie de la chane alimentaire - Validation des mthodes - Partie 2: Protocole pour la validation de mthodes alternat
14、ives (commerciales) par rapport une mthode de rfrence (ISO 16140-2:2016) Mikrobiologie der Lebensmittelkette - Verfahrensvalidierung - Teil 2: Arbeitsvorschrift fr die Validierung von alternativen (urheberrechtlich geschtzten) Verfahren anhand eines Referenzverfahrens (ISO 16140-2:2016) This Europea
15、n Standard was approved by CEN on 12 May 2016. 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 concerni
16、ng 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 i
17、ts 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, Former Yugoslav Republic of Macedonia, France, Germ
18、any, Greece, Hungary, Iceland, Ireland, Italy, Latvia, Lithuania, Luxembourg, Malta, Netherlands, Norway, Poland, Portugal, Romania, Slovakia, Slovenia, Spain, Sweden, Switzerland, Turkey and United Kingdom. EUROPEAN COMMITTEE FOR STANDARDIZATION COMIT EUROPEN DE NORMALISATION EUROPISCHES KOMITEE FR
19、 NORMUNG CEN-CENELEC Management Centre: Avenue Marnix 17, B-1000 Brussels 2016 CEN All rights of exploitation in any form and by any means reserved worldwide for CEN national Members. Ref. No. EN ISO 16140-2:2016 EEuropean foreword .3Introduction 51 Scope . 62 Normative references 63 Terms and defin
20、itions . 64 General principles for the validation of alternative methods . 65 Qualitative methods Technical protocol for validation 75.1 Method comparison study 75.1.1 General considerations . 75.1.2 Paired or unpaired study 75.1.3 Sensitivity study . 75.1.4 Relative level of detection study 125.1.5
21、 Inclusivity and exclusivity study . 145.2 Interlaboratory study . 155.2.1 General considerations 155.2.2 Measurement protocol 155.2.3 Calculations and summary of data 175.2.4 Interpretation of data .206 Quantitative methods Technical protocol for validation .216.1 Method comparison study . 216.1.1
22、General considerations 216.1.2 Relative trueness study . 216.1.3 Accuracy profile study256.1.4 Limit of quantification study 296.1.5 Inclusivity and exclusivity study . 296.2 Interlaboratory study . 316.2.1 General considerations 316.2.2 Measurement protocol 316.2.3 Calculations, summary, and interp
23、retation of data .32Annex A (informative) Classification of sample types and suggested target combinations for validation studies .35Annex B (normative) Order of preference for use of naturally and artificially contaminated samples in validation studies .51Annex C (informative) General protocols for
24、 contamination by mixture and artificial contamination of foods .52Annex D (informative) Models for RLOD calculations using data from the method comparison study .55Annex E (normative) Points to be considered when selecting strains for testing inclusivity and exclusivity .57Annex F (informative) Con
25、siderations for calculations of the relative level of detection (RLOD) between laboratories as obtained in an interlaboratory study 59Annex G (informative) Principle of the accuracy profile for validation of quantitative models .62Annex H (informative) Application of the accuracy profile in the meth
26、od comparison study 64Annex I (informative) Example of the application of the accuracy profile for an interlaboratory study .67Bibliography .71Contents PageDIN EN ISO 16140-2:2016-11 EN ISO 16140-2:2016 (E) 2Foreword 4European foreword This document (EN ISO 16140-2:2016) has been prepared by Technic
27、al Committee ISO/TC 34 “Food products“ in collaboration with Technical Committee CEN/TC 275 “Food analysis - Horizontal methods” the secretariat of which is held by DIN. This European Standard shall be given the status of a national standard, either by publication of an identical text or by endorsem
28、ent, at the latest by January 2017, and conflicting national standards shall be withdrawn at the latest by January 2017. 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 identify
29、ing any or all such patent rights. This document supersedes EN ISO 16140:2003. 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
30、, Denmark, Estonia, Finland, Former Yugoslav Republic of Macedonia, France, Germany, Greece, Hungary, Iceland, Ireland, Italy, Latvia, Lithuania, Luxembourg, Malta, Netherlands, Norway, Poland, Portugal, Romania, Slovakia, Slovenia, Spain, Sweden, Switzerland, Turkey and the United Kingdom. Endorsem
31、ent notice The text of ISO 16140-2:2016 has been approved by CEN as EN ISO 16140-2:2016 without any modification. DIN EN ISO 16140-2:2016-11 EN ISO 16140-2:2016 (E) 3 ForewordISO (the International Organization for Standardization) is a worldwide federation of national standards bodies (ISO member b
32、odies). 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 a
33、nd 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.The procedures used to develop this document and those intended for its further maintenance are d
34、escribed in the ISO/IEC Directives, Part 1. In particular the different approval criteria needed for the different types of ISO documents should be noted. This document was drafted in accordance with the editorial rules of the ISO/IEC Directives, Part 2 (see www.iso.org/directives).Attention is draw
35、n 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. Details of any patent rights identified during the development of the document will be in the Introduction and/or on t
36、he ISO list of patent declarations received (see www.iso.org/patents).Any trade name used in this document is information given for the convenience of users and does not constitute an endorsement.For an explanation on the meaning of ISO specific terms and expressions related to conformity assessment
37、, as well as information about ISOs adherence to the WTO principles in the Technical Barriers to Trade (TBT) see the following URL: Foreword - Supplementary Information The committee responsible for this document is ISO/TC 34, Food products, Subcommittee SC 9, Microbiology.This first edition of ISO
38、16140-2, together with ISO 16140-1, cancels and replaces ISO 16140:2003, which has been technically revised. It also incorporates the Amendment ISO 16140:2003:Amd.1:2011.ISO 16140 consists of the following parts, under the general title Microbiology of the food chain Method validation: Part 1: Vocab
39、ulary Part 2: Protocol for the validation of alternative (proprietary) methods against a reference methodThe following parts are under preparation: Part 3: Protocol for the verification of reference and validated alternative methods implemented in a single laboratory Part 4: Protocol for single-labo
40、ratory (in-house) method validation Part 5: Protocol for factorial interlaboratory validation of non-proprietary methods Part 6: Protocol for the validation of alternative (proprietary) methods for microbiological confirmation and typing DIN EN ISO 16140-2:2016-11 EN ISO 16140-2:2016 (E) 4 Introduct
41、ionToday, many alternative, mostly proprietary, methods exist that are used to assess the microbiological quality of raw materials and finished products and the microbiological status of manufacturing procedures. These methods are often faster and easier to perform than the corresponding standardize
42、d method. The developers, end users, and authorities need a reliable common protocol for the validation of such alternative methods. The data generated will also provide potential end users with performance data for a given method, thus, enabling them to make an informed choice on the adoption of a
43、particular method. The data generated can also be the basis for the certification of a method by an independent organization.This part of ISO 16140 is intended to provide a specific protocol and guidelines for the validation of proprietary methods intended to be used as a rapid and/or easier method
44、to perform than the corresponding reference method, can also be used for the validation of other non-proprietary methods that are used instead of the reference method, is intended as the successor of the validation protocol published in the first version of ISO 16140 (ISO 16140:2003), and is mainly
45、written for the validation of methods that are capable of culturing the target microorganism, but can also be applied to methods for microorganisms that cannot be cultured such as viruses (e.g. Norovirus) and protozan parasites (e.g. Cryptosporidium or Giardia). In these cases, some wordings are to
46、be interpreted so as to fit the situation for non-culturable organisms.The use of this part of ISO 16140 involves expertise on relevant areas such as microbiology, statistical design, and analysis as indicated in the respective sections. The statistical expertise encompasses overview of sampling the
47、ory and design of experiments, statistical analysis of (qualitative and quantitative) microbiological data, and overview of statistical concepts on random sampling, sample heterogeneity, sample stability, design of experiments, and variance components.When this part of ISO 16140 is next reviewed, ac
48、count will be taken of all information then available regarding the extent to which the guidelines have been followed and the reasons for deviation from them in the case of particular products.The harmonization of validation methods cannot be immediate and for certain groups of products, International Standards and/or national standards may already exist that do not comply with this part of ISO 16140. It is hoped that when such standards are reviewed, they will be changed to comply with ISO 16140 so that eventually, the only remaining departures from this part of ISO 16
