1、BS EN ISO 16140-2:2016Microbiology of the food chain Method validationPart 2: Protocol for the validation of alternative (proprietary) methods against a reference method (ISO 16140-2:2016)BSI Standards PublicationWB11885_BSI_StandardCovs_2013_AW.indd 1 15/05/2013 15:06BS EN ISO 16140-2:2016 BRITISH
2、STANDARDNational forewordThis British Standard is the UK implementation of EN ISO16140-2:2016. Together with BS EN ISO 16140-1:2016, it supersedes BS EN ISO 16140:2003+A1:2011 which is withdrawn.The UK participation in its preparation was entrusted to TechnicalCommittee AW/9, Microbiology.A list of
3、organizations represented on this committee can beobtained on request to its secretary.This publication does not purport to include all the necessaryprovisions of a contract. Users are responsible for its correctapplication. The British Standards Institution 2016.Published by BSI Standards Limited 2
4、016ISBN 978 0 580 69940 5ICS 07.100.30Compliance with a British Standard cannot confer immunity fromlegal obligations.This British Standard was published under the authority of theStandards Policy and Strategy Committee on 31 July 2016.Amendments/corrigenda issued since publicationDate T e x t a f f
5、 e c t e dEUROPEAN STANDARD NORME EUROPENNE EUROPISCHE NORM EN ISO 16140-2 July 2016 ICS 07.100.30 Supersedes EN ISO 16140:2003English Version Microbiology of the food chain - Method validation - Part 2: Protocol for the validation of alternative (proprietary) methods against a reference method (ISO
6、 16140-2:2016)Microbiologie de la chane alimentaire - Validation des mthodes - Partie 2: Protocole pour la validation de mthodes alternatives (commerciales) par rapport une mthode de rfrence (ISO 16140-2:2016) Mikrobiologie der Lebensmittelkette - Verfahrensvalidierung - Teil 2: Arbeitsvorschrift fr
7、 die Validierung von alternativen (urheberrechtlich geschtzten) Verfahren anhand eines Referenzverfahrens (ISO 16140-2:2016) This European 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
8、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 officia
9、l 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 Au
10、stria, Belgium, Bulgaria, Croatia, Cyprus, Czech Republic, 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, Swe
11、den, Switzerland, Turkey and United Kingdom. EUROPEAN COMMITTEE FOR STANDARDIZATION COMIT EUROPEN DE NORMALISATION EUROPISCHES KOMITEE FR 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 C
12、EN national Members. Ref. No. EN ISO 16140-2:2016 EBS EN ISO 16140-2:2016EN ISO 16140-2:2016 (E) 3 European foreword This document (EN ISO 16140-2:2016) has been prepared by Technical Committee ISO/TC 34 “Food products“ in collaboration with Technical Committee CEN/TC 275 “Food analysis - Horizontal
13、 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 endorsement, at the latest by January 2017, and conflicting national standards shall be withdrawn at the latest by January 2017
14、. 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. This document supersedes EN ISO 16140:2003. According to the CEN-CENELEC Internal Re
15、gulations, 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, Former Yugoslav Republic of Macedonia, France, Germany, Greece, Hungary, Iceland, Ireland,
16、 Italy, Latvia, Lithuania, Luxembourg, Malta, Netherlands, Norway, Poland, Portugal, Romania, Slovakia, Slovenia, Spain, Sweden, Switzerland, Turkey and the United Kingdom. Endorsement notice The text of ISO 16140-2:2016 has been approved by CEN as EN ISO 16140-2:2016 without any modification. BS EN
17、 ISO 16140-2:2016ISO 16140-2:2016(E)Foreword ivIntroduction v1 Scope . 12 Normative references 13 Terms and definitions . 14 General principles for the validation of alternative methods . 15 Qualitative methods Technical protocol for validation 25.1 Method comparison study 25.1.1 General considerati
18、ons . 25.1.2 Paired or unpaired study 25.1.3 Sensitivity study . 25.1.4 Relative level of detection study . 75.1.5 Inclusivity and exclusivity study 95.2 Interlaboratory study . 105.2.1 General considerations 105.2.2 Measurement protocol 105.2.3 Calculations and summary of data 125.2.4 Interpretatio
19、n of data .156 Quantitative methods Technical protocol for validation .166.1 Method comparison study . 166.1.1 General considerations 166.1.2 Relative trueness study . 166.1.3 Accuracy profile study206.1.4 Limit of quantification study 246.1.5 Inclusivity and exclusivity study . 246.2 Interlaborator
20、y study . 266.2.1 General considerations 266.2.2 Measurement protocol 266.2.3 Calculations, summary, and interpretation of data .27Annex A (informative) Classification of sample types and suggested target combinations for validation studies .30Annex B (normative) Order of preference for use of natur
21、ally and artificially contaminated samples in validation studies .46Annex C (informative) General protocols for contamination by mixture and artificial contamination of foods .47Annex D (informative) Models for RLOD calculations using data from the method comparison study .50Annex E (normative) Poin
22、ts to be considered when selecting strains for testing inclusivity and exclusivity .52Annex F (informative) Considerations for calculations of the relative level of detection (RLOD) between laboratories as obtained in an interlaboratory study 54Annex G (informative) Principle of the accuracy profile
23、 for validation of quantitative models .57Annex H (informative) Application of the accuracy profile in the method comparison study 59Annex I (informative) Example of the application of the accuracy profile for an interlaboratory study .62Bibliography .66 ISO 2016 All rights reserved iiiContents Page
24、BS EN ISO 16140-2:2016ISO 16140-2:2016(E)ForewordISO (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 bo
25、dy 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 Electrot
26、echnical Commission (IEC) on all matters of electrotechnical standardization.The procedures used to develop this document and those intended for its further maintenance are described in the ISO/IEC Directives, Part 1. In particular the different approval criteria needed for the different types of IS
27、O 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 drawn to the possibility that some of the elements of this document may be the subject of patent rights. ISO shall not be held res
28、ponsible 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 the ISO list of patent declarations received (see www.iso.org/patents).Any trade name used in this document is information give
29、n 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, as well as information about ISOs adherence to the WTO principles in the Technical Barriers to Trade (TBT) see the following
30、 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 16140-2, together with ISO 16140-1, cancels and replaces ISO 16140:2003, which has been technically revised. It also incorpora
31、tes 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: Vocabulary Part 2: Protocol for the validation of alternative (proprietary) methods against a reference methodThe following parts a
32、re under preparation: Part 3: Protocol for the verification of reference and validated alternative methods implemented in a single laboratory Part 4: Protocol for single-laboratory (in-house) method validation Part 5: Protocol for factorial interlaboratory validation of non-proprietary methods Part
33、6: Protocol for the validation of alternative (proprietary) methods for microbiological confirmation and typing iv ISO 2016 All rights reservedBS EN ISO 16140-2:2016ISO 16140-2:2016(E)IntroductionToday, many alternative, mostly proprietary, methods exist that are used to assess the microbiological q
34、uality 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 standardized method. The developers, end users, and authorities need a reliable common protocol for the validation of
35、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 particular method. The data generated can also be the basis for the certification of a method by an indepen
36、dent 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 to perform than the corresponding reference method, can also be used for the validation of other non-propri
37、etary 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 written for the validation of methods that are capable of culturing the target microorganism, but can also
38、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 be interpreted so as to fit the situation for non-culturable organisms.The use of this part of ISO 16140 in
39、volves expertise on relevant areas such as microbiology, statistical design, and analysis as indicated in the respective sections. The statistical expertise encompasses overview of sampling theory and design of experiments, statistical analysis of (qualitative and quantitative) microbiological data,
40、 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, account will be taken of all information then available regarding the extent to which the guidelines have bee
41、n 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
42、 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 16140 will be those necessary for well-established technical reasons. For example, ISO 162973deals with a very specific validation fo
43、r a specific subject (the hygienic status of raw milk samples) and will remain as a vertical standard besides ISO 16140. If such a validation is needed, the vertical standard is more important. ISO 2016 All rights reserved vBS EN ISO 16140-2:2016BS EN ISO 16140-2:2016Microbiology of the food chain M
44、ethod validation Part 2: Protocol for the validation of alternative (proprietary) methods against a reference method1 ScopeThis part of ISO 16140 specifies the general principle and the technical protocol for the validation of alternative, mostly proprietary, methods for microbiology in the food cha
45、in. Validation studies according to this part of ISO 16140 are intended to be performed by organizations involved in method validation.This part of ISO 16140 is applicable to the validation of methods for the analysis (detection or quantification) of microorganisms in products intended for human con
46、sumption, products intended for animal feeding, environmental samples in the area of food and feed production, handling, and samples from the primary production stage.This part of ISO 16140 is in particular applicable to bacteria and fungi. Some clauses of this part of ISO 16140 could be applicable
47、to other (micro) organisms or their metabolites on a case-by-case-basis. In the future, guidance for other organisms (e.g. viruses and parasites) will be included in either this part or a separate part of ISO 16140.2 Normative referencesThe following documents, in whole or in part, are normatively r
48、eferenced in this document and are indispensable for its application. For dated references, only the edition cited applies. For undated references, the latest edition of the referenced document (including any amendments) applies.ISO 16140-1, Microbiology of the food chain Method validation Part 1: V
49、ocabulary3 Terms and definitionsFor the purposes of this document, the terms and definitions given in ISO 16140-1 apply.4 General principles for the validation of alternative methodsThe validation protocol comprises two phases: a method comparison study of the alternative (proprietary) method against the reference method carried out in the organizing laboratory; an interlaboratory study of the alternative (proprietary) method against the reference method carried out in different laboratories.The technical rules for performing the
