EN ISO 17468:2023

SLOVENSKI STANDARD
01-marec-2024
Nadomešča:
SIST EN ISO 17468:2016
Mikrobiologija v prehranski verigi - Tehnične zahteve in navodila za vzpostavitev
ali revizijo standardnih referenčnih metod (ISO 17468:2023)
Microbiology of the food chain - Technical requirements and guidance on the
establishment or revision of a standardized reference method (ISO 17468:2023)
Mikrobiologie der Lebensmittelkette - Technische Anforderungen und Leitfaden zur
Einführung oder Überarbeitung von genormten Referenzverfahren (ISO 17468:2023)
Microbiologie de la chaîne alimentaire - Exigences et recommandations techniques pour
le développement ou la révision d'une méthode de référence normalisée (ISO
17468:2023)
Ta slovenski standard je istoveten z: EN ISO 17468:2023
ICS:
07.100.30 Mikrobiologija živil Food microbiology
2003-01.Slovenski inštitut za standardizacijo. Razmnoževanje celote ali delov tega standarda ni dovoljeno.

EN ISO 17468
EUROPEAN STANDARD
NORME EUROPÉENNE
November 2023
EUROPÄISCHE NORM
ICS 07.100.30 Supersedes EN ISO 17468:2016
English Version
Microbiology of the food chain - Technical requirements
and guidance on the establishment or revision of a
standardized reference method (ISO 17468:2023)
Microbiologie de la chaîne alimentaire - Exigences et Mikrobiologie der Lebensmittelkette - Technische
recommandations techniques pour le développement Anforderungen und Leitfaden zur Einführung oder
ou la révision d'une méthode de référence normalisée Überarbeitung von genormten Referenzverfahren (ISO
(ISO 17468:2023) 17468:2023)
This European Standard was approved by CEN on 29 September 2023.

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
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 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 national 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, Republic of North Macedonia, Romania, Serbia, Slovakia, Slovenia, Spain, Sweden, Switzerland, Türkiye and
United Kingdom.
EUROPEAN COMMITTEE FOR STANDARDIZATION
COMITÉ EUROPÉEN DE NORMALISATION

EUROPÄISCHES KOMITEE FÜR NORMUNG

CEN-CENELEC Management Centre: Rue de la Science 23, B-1040 Brussels
© 2023 CEN All rights of exploitation in any form and by any means reserved Ref. No. EN ISO 17468:2023 E
worldwide for CEN national Members.

Contents Page
European foreword . 3

European foreword
This document (EN ISO 17468:2023) has been prepared by Technical Committee ISO/TC 34 "Food
products" in collaboration with Technical Committee CEN/TC 463 “Microbiology of the food chain” the
secretariat of which is held by AFNOR.
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 May 2024, and conflicting national standards shall be
withdrawn at the latest by May 2024.
Attention is drawn to the possibility that some of the elements of this document may be the subject of
patent rights. CEN shall not be held responsible for identifying any or all such patent rights.
This document supersedes EN ISO 17468:2016.
Any feedback and questions on this document should be directed to the users’ national standards
body/national committee. A complete listing of these bodies can be found on the CEN website.
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, Denmark, Estonia, Finland, France, Germany, Greece, Hungary, Iceland,
Ireland, Italy, Latvia, Lithuania, Luxembourg, Malta, Netherlands, Norway, Poland, Portugal, Republic of
North Macedonia, Romania, Serbia, Slovakia, Slovenia, Spain, Sweden, Switzerland, Türkiye and the
United Kingdom.
Endorsement notice
The text of ISO 17468:2023 has been approved by CEN as EN ISO 17468:2023 without any modification.

INTERNATIONAL ISO
STANDARD 17468
Second edition
2023-11
Microbiology of the food chain —
Technical requirements and guidance
on the establishment or revision of a
standardized reference method
Microbiologie de la chaîne alimentaire — Exigences et
recommandations techniques pour le développement ou la révision
d'une méthode de référence normalisée
Reference number
ISO 17468:2023(E)
ISO 17468:2023(E)
© ISO 2023
All rights reserved. Unless otherwise specified, or required in the context of its implementation, no part of this publication may
be reproduced or utilized otherwise in any form or by any means, electronic or mechanical, including photocopying, or posting on
the internet or an intranet, without prior written permission. Permission can be requested from either ISO at the address below
or ISO’s member body in the country of the requester.
ISO copyright office
CP 401 • Ch. de Blandonnet 8
CH-1214 Vernier, Geneva
Phone: +41 22 749 01 11
Email: copyright@iso.org
Website: www.iso.org
Published in Switzerland
ii
ISO 17468:2023(E)
Contents Page
Foreword .iv
1 Scope . 1
2 Normative references . 1
3 Terms and definitions . 1
4 Technical procedure for standardizing a new reference method.3
4.1 General . 3
4.2 Technical steps . 3
4.2.1 Step 1: Method(s) selection (mandatory) . 3
4.2.2 Step 2: Method(s) optimization (optional) . 4
4.2.3 Step 3: Method(s) evaluation study (recommended) . 4
4.2.4 Step 4: “Real life” study (recommended) . 5
4.2.5 Step 5: Selection of one candidate reference method for further validation
(mandatory) . 5
4.2.6 Step 6: Interlaboratory study (mandatory) . 5
5 Technical procedure for revising a standardized reference method .7
Annex A (informative) Flow chart on technical steps for the establishment or revision of a
standardized reference method.9
Annex B (informative) Guidance on optimization studies to compare two options in the
development of a new standardized reference method or for its revision .10
Bibliography .13
iii
ISO 17468:2023(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 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 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.
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 ISO document 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).
ISO draws attention to the possibility that the implementation of this document may involve the use
of (a) patent(s). ISO takes no position concerning the evidence, validity or applicability of any claimed
patent rights in respect thereof. As of the date of publication of this document, ISO had not received
notice of (a) patent(s) which may be required to implement this document. However, implementers are
cautioned that this may not represent the latest information, which may be obtained from the patent
database available at www.iso.org/patents. ISO shall not be held responsible for identifying any or all
such patent rights.
Any trade name used in this document is information given for the convenience of users and does not
constitute an endorsement.
For an explanation of the voluntary nature of standards, the meaning of ISO specific terms and
expressions related to conformity assessment, as well as information about ISO's adherence to
the World Trade Organization (WTO) principles in the Technical Barriers to Trade (TBT), see
www.iso.org/iso/foreword.html.
This document was prepared by Technical Committee ISO/TC 34, Food products, Subcommittee SC 9,
Microbiology, in collaboration with the European Committee for Standardization (CEN) Technical
Committee CEN/TC 463, Microbiology of the food chain, in accordance with the Agreement on technical
cooperation between ISO and CEN (Vienna Agreement).
This second edition cancels and replaces the first edition (ISO 17468:2016), which has been technically
revised.
The main changes are as follows:
— a cross-reference is made not only to ISO 16140-2, but also to ISO 16140-4 and ISO 16140-6;
— a new optional step has been added, "method(s) optimization". In addition, a new annex providing
guidance on method optimization studies is included, to compare two options during the development
of a new standardized reference method or for its revision;
— the inclusion of the case of confirmation and typing methods;
— the assessment of the nature of a change (minor/major) during the revision of a standardized
reference method.
Any feedback or questions on this document should be directed to the user’s national standards body. A
complete listing of these bodies can be found at www.iso.org/members.html.
iv
INTERNATIONAL STANDARD ISO 17468:2023(E)
Microbiology of the food chain — Technical requirements
and guidance on the establishment or revision of a
standardized reference method
1 Scope
This document gives technical requirements and guidance on the establishment or revision of
standardized reference methods used for the analysis of microorganisms in:
— products intended for human consumption;
— products for feeding animals;
— environmental samples in the area of food and feed production and handling;
— samples from the primary production stage.
This document specifies the technical stages of the establishment of a new standardized reference
method and of the revision of an existing standardized reference method. It includes, in particular,
requirements and guidance on the validation of the selected method.
This document is intended to be implemented in particular by ISO/TC 34/SC 9 and its corresponding
structure at CEN level, which is CEN/TC 463.
2 Normative references
The following documents are referred to in the text in such a way that some or all of their content
constitutes requirements of this document. For dated references, only the edition cited applies. For
undated references, the latest edition of the referenced document (including any amendments) applies.
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
ISO 11133, Microbiology of food, animal feed and water — Preparation, production, storage and
performance testing of culture media
ISO 16140-1:2016, Microbiology of the food chain — Method validation — Part 1: Vocabulary
ISO 16140-2:2016, Microbiology of the food chain — Method validation — Part 2: Protocol for the validation
of alternative (proprietary) methods against a reference method
1)
ISO 16140-2:2016/Amd.1:— , Microbiology of the food chain — Method validation — Part 2: Protocol for
the validation of alternative (proprietary) methods against a reference method — Amendment 1: Revision
of qualitative MCS data evaluation, RLOD calculations in the ILS, calculation and interpretation of the RT
study, and inclusion of a commercial sterility testing protocol for specific products
ISO 16140-6:2019, Microbiology of the food chain — Method validation — Part 6: Protocol for the validation
of alternative (proprietary) methods for microbiological confirmation and typing procedures
3 Terms and definitions
For the purposes of this document, the terms and definitions given in ISO 16140-1 and the following
apply.
1) Under preparation. Stage at time of publication ISO 16140-2:2016/DAmd.1:2023.
ISO 17468:2023(E)
ISO and IEC maintain terminology databases for use in standardization at the following addresses:
— ISO Online browsing platform: available at https:// www .iso .org/ obp
— IEC Electropedia: available at https:// www .electropedia .org/
3.1
candidate reference method
method selected and likely to become the standardized reference method (3.7)
3.2
ILS organizer
organizing laboratory
laboratory with responsibility for managing all of the technical and statistical activities involved in the
organization of the interlaboratory study
[SOURCE: ISO 16140-1:2016, 2.45, modified - “validation study, i.e. method comparison study and the
interlaboratory study” has been replaced by “organization of the interlaboratory study” and the Note 1
to entry has been deleted.]
3.3
ILS participant
participating laboratory
individual laboratory technician, who works completely independently from other ILS participants,
using different sets of blind samples or test portions
[SOURCE: ISO 16140-1:2016, 2.13, modified - “collaborator” has been replaced by “ILS participant”.]
3.4
interlaboratory study
study performed by multiple laboratories testing identical samples at the same time, the results of
which are used to estimate performance characteristics of the candidate reference method
Note 1 to entry: The aim of an interlaboratory study is to determine the variability of the results obtained in
different laboratories using identical samples.
[SOURCE: ISO 16140-1:2016, 2.33, modified - “alternative-method performance parameters” has been
replaced by “performance characteristics of the candidate reference method”.]
3.5
pre-standardization stage
technical stage prior to the standardization stage and comprising the different steps described in this
document
Note 1 to entry: The standardization stage starts with the proposal stage which is the approval of a New Work
Item Proposal (ISO/NP) for inclusion of the Work Item on the work programme of ISO/TC 34/SC 9.
3.6
“real life” study
study of one or several methods, conducted in different laboratories, using their own routine samples
and with preference given to naturally contaminated samples
3.7
standardized reference method
reference method described in a standard
Note 1 to entry: See ISO 16140-1 for the definition of “reference method”.
ISO 17468:2023(E)
4 Technical procedure for standardizing a new reference method
4.1 General
The validation of a method in view of its standardization as a reference method includes six technical
steps (see 4.2):
— step 1: method(s) selection (mandatory);
— step 2: method(s) optimization (optional);
— step 3: method(s) evaluation study (recommended);
— step 4: “real life” study (recommended) (this step does not apply to confirmation and typing
methods);
— step 5: selection of one candidate reference method for further validation (mandatory);
— step 6: interlaboratory study (mandatory).
The first five technical steps correspond to a pre-standardization stage and are usually performed
before launching the standardization process. Step 6 (see 4.2.6) is usually performed during the
standardization process [preferably after the committee stage (ISO/CD) and before the enquiry stage
(ISO/DIS)].
The flow chart of the technical steps for the establishment of a new standardized reference method is
given in Annex A.
The working group in charge of organizing the studies on a qualitative method shall consider the
inclusion of a test portion size larger than, for example, 10 g or 25 g, when it is relevant and feasible.
This will facilitate the use of the standardized reference method without the need for further validation
when a larger test portion size is routinely used.
EXAMPLE Detection of Salmonella in 375 g infant formula test portions.
Information on the studies conducted in step 1 to step 6 should not be shared publicly until their
completion. Once these studies are completed, all relevant data as obtained in step 1 to step 6 should
become publicly available, by reporting it either in a scientific publication or in a report. The report
associated to each standardized reference method can be made available on the ISO Standards
Maintenance Portal (https:// standards .iso .org/ iso/ ) and/or by including a link in the Bibliography of
the standardized reference method to the website where the report is publicly available.
NOTE Data, e.g. regarding inclusivity and exclusivity, can be derived from earlier studies as long as the
information is traceable to the originally published data or made available on the ISO Standards Maintenance
Portal associated to the standardized reference method.
The performance characteristics obtained from the interlaboratory study (step 6, also see 4.2.6) shall
be incorporated into the corresponding standardized reference method.
4.2 Technical steps
4.2.1 Step 1: Method(s) selection (mandatory)
Information from different sources (e.g. national/regional standardized methods, scientific papers on
methods with evaluation data, evaluation/validation reports on methods, practicability of the method)
shall be collated for the choice of (a) candidate reference method(s). Based on the information available,
the working group in charge of developing the standard (referred to as ‘the working group’ from this
point forward) shall select one or several candidate reference methods.
ISO 17468:2023(E)
4.2.2 Step 2: Method(s) optimization (optional)
If, after the step of method(s) selection (see 4.2.1), one (or several) candidate reference method(s)
comprise(s) two options for one factor (e.g. culture medium, incubation time, incubation temperature),
a method(s) optimization study can be conducted, comparing the two options.
Annex B provides guidance on how to compare these two options, and how to make a choice for the
method to be standardized and to be further evaluated (see 4.2.3).
If the candidate reference method(s) does (do) not comprise(s) any factor with options, proceed directly
to step 3 (see 4.2.3).
4.2.3 Step 3: Method(s) evaluation study (recommended)
4.2.3.1 General
An evaluation study of the candidate reference method(s) (see 4.2.1 and 4.2.2) should be conducted,
normally by one
...