SIST EN ISO 16140-3:2021/A1:2025
(Amendment)Microbiology of the food chain - Method validation - Part 3: Protocol for the verification of reference methods and validated alternative methods in a single laboratory - Amendment 1: Protocol for verification of validated identification methods of microorganisms (ISO 16140-3:2021/Amd 1:2025)
Microbiology of the food chain - Method validation - Part 3: Protocol for the verification of reference methods and validated alternative methods in a single laboratory - Amendment 1: Protocol for verification of validated identification methods of microorganisms (ISO 16140-3:2021/Amd 1:2025)
Mikrobiologie der Lebensmittelkette - Verfahrensvalidierung - Teil 3: Arbeitsvorschrift für die Verifizierung von Referenz- und validierten alternativen Verfahren in einem Einzel-Labor - Änderung 1: Verifizierung von validierten Identifizierungsverfahren (ISO 16140-3:2021/Amd 1:2025)
Microbiologie de la chaîne alimentaire - Validation des méthodes - Partie 3: Protocole pour la vérification dans un seul laboratoire de méthodes de référence et de méthodes alternatives validées - Amendement 1: Protocole pour la vérification des méthodes validées d’identification des micro-organismes (ISO 16140-3:2021/Amd 1:2025)
Mikrobiologija v prehranski verigi - Validacija metode - 3. del: Protokol za preverjanje referenčnih in validiranih alternativnih metod, izvedenih v posameznem laboratoriju - Dopolnilo A1: Protokol za preverjanje validiranih metod za identifikacijo mikroorganizmov (ISO 16140-3:2021/Amd1:2025)
General Information
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Standards Content (Sample)
SLOVENSKI STANDARD
01-november-2025
Mikrobiologija v prehranski verigi - Validacija metode - 3. del: Protokol za
preverjanje referenčnih in validiranih alternativnih metod, izvedenih v
posameznem laboratoriju - Dopolnilo A1: Protokol za preverjanje validiranih metod
za identifikacijo mikroorganizmov (ISO 16140-3:2021/Amd1:2025)
Microbiology of the food chain - Method validation - Part 3: Protocol for the verification of
reference methods and validated alternative methods in a single laboratory - Amendment
1: Protocol for verification of validated identification methods of microorganisms (ISO
16140-3:2021/Amd 1:2025)
Mikrobiologie der Lebensmittelkette - Verfahrensvalidierung - Teil 3: Arbeitsvorschrift für
die Verifizierung von Referenz- und validierten alternativen Verfahren in einem Einzel-
Labor - Änderung 1: Verifizierung von validierten Identifizierungsverfahren (ISO 16140-
3:2021/Amd 1:2025)
Microbiologie de la chaîne alimentaire - Validation des méthodes - Partie 3: Protocole
pour la vérification dans un seul laboratoire de méthodes de référence et de méthodes
alternatives validées - Amendement 1: Protocole pour la vérification des méthodes
validées d’identification des micro-organismes (ISO 16140-3:2021/Amd 1:2025)
Ta slovenski standard je istoveten z: EN ISO 16140-3:2021/A1:2025
ICS:
07.100.30 Mikrobiologija živil Food microbiology
2003-01.Slovenski inštitut za standardizacijo. Razmnoževanje celote ali delov tega standarda ni dovoljeno.
EN ISO 16140-3:2021/A1
EUROPEAN STANDARD
NORME EUROPÉENNE
August 2025
EUROPÄISCHE NORM
ICS 07.100.30
English Version
Microbiology of the food chain - Method validation - Part 3:
Protocol for the verification of reference methods and
validated alternative methods in a single laboratory -
Amendment 1: Protocol for verification of validated
identification methods of microorganisms (ISO 16140-
3:2021/Amd 1:2025)
Microbiologie de la chaîne alimentaire - Validation des Mikrobiologie der Lebensmittelkette -
méthodes - Partie 3: Protocole pour la vérification dans Verfahrensvalidierung - Teil 3: Arbeitsvorschrift für die
un seul laboratoire de méthodes de référence et de Verifizierung von Referenz- und validierten
méthodes alternatives validées - Amendement 1: alternativen Verfahren in einem Einzel-Labor -
Protocole pour la vérification des méthodes validées Änderung 1: Validierte Identifizierungsverfahren von
d'identification des micro-organismes (ISO 16140- Mikroorganismen - Technische Arbeitsvorschrift zur
3:2021/Amd 1:2025) Verifizierung (ISO 16140-3:2021/Amd 1:2025)
This amendment A1 modifies the European Standard EN ISO 16140-3:2021; it was approved by CEN on 25 August 2025.
CEN members are bound to comply with the CEN/CENELEC Internal Regulations which stipulate the conditions for inclusion of
this amendment into the relevant 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 amendment 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
© 2025 CEN All rights of exploitation in any form and by any means reserved Ref. No. EN ISO 16140-3:2021/A1:2025 E
worldwide for CEN national Members.
Contents Page
European foreword . 3
European foreword
This document (EN ISO 16140-3:2021/A1:2025) 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 Amendment to the European Standard EN ISO 16140-3:2021 shall be given the status of a national
standard, either by publication of an identical text or by endorsement, at the latest by February 2026,
and conflicting national standards shall be withdrawn at the latest by February 2026.
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.
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 16140-3:2021/Amd 1:2025 has been approved by CEN as EN ISO 16140-
3:2021/A1:2025 without any modification.
International
Standard
ISO 16140-3
First edition
Microbiology of the food chain —
2021-01
Method validation —
AMENDMENT 1
Part 3:
2025-08
Protocol for the verification of
reference methods and validated
alternative methods in a single
laboratory
AMENDMENT 1: Protocol for
verification of validated identification
methods of microorganisms
Microbiologie de la chaîne alimentaire — Validation des
méthodes —
Partie 3: Protocole pour la vérification dans un seul laboratoire
de méthodes de référence et de méthodes alternatives validées
AMENDEMENT 1: Protocole pour la vérification des méthodes
validées d’identification des micro-organismes
Reference number
ISO 16140-3:2021/Amd.1:2025(en) © ISO 2025
ISO 16140-3:2021/Amd.1:2025(en)
© ISO 2025
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
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Email: copyright@iso.org
Website: www.iso.org
Published in Switzerland
ii
ISO 16140-3:2021/Amd.1:2025(en)
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).
A list of all parts in the ISO 16140 series can be found on the ISO website.
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.
iii
ISO 16140-3:2021/Amd.1:2025(en)
Microbiology of the food chain — Method validation —
Part 3:
Protocol for the verification of reference methods and
validated alternative methods in a single laboratory
AMENDMENT 1: Protocol for verification of validated
identification methods of microorganisms
Introduction (0.1), first paragraph
Replace the third sentence with:
The ISO 16140 series consists of several parts with the general title, Microbiology of the food chain — Method
validation:
Add the following text after the sixth list item:
— Part 7: Protocol for the validation of identification methods of microorganisms.
Introduction (0.1), sixth paragraph
Replace the reference to ISO 17468:2016, 3.5, with:
In the ISO 16140 series, reference methods include standardized reference (ISO and CEN) methods as
defined in ISO 17468:2023, 3.7, as a “reference method described in a standard”.
Introduction (0.1), tenth paragraph
Replace the first sentence with:
ISO 16140-6 and ISO 16140-7 are somewhat different from the other parts in the ISO 16140 series in that
they relate to very specific situations. ISO 16140-6 is restricted to the confirmation procedure of a method
to be validated [e.g. the biochemical confirmation of Enterobacteriaceae (see ISO 21528-2)].
Replace the verb “defines” with “specifies” in the fourth sentence:
The validation study in ISO 16140-6 clearly specifies the selective agar(s) from which strains can be
confirmed using the alternative confirmation method.
ISO 16140-3:2021/Amd.1:2025(en)
Introduction (0.1), Figure 2
Replace Figure 2 with:
Figure 2 — Use of validated alternative confirmation methods (see ISO 16140-6)
Introduction (0.1), Example
Number the example to Example 1:
EXAMPLE 1 An example application of a validated alternative confirmation method is as follows:
Introduction (0.1), after the last paragraph
Add the following text in the end of 0.1:
ISO 16140-7 addresses the validation of identification procedures (e.g. molecular identification using
multiplex PCR or DNA sequencing or mass spectrometry). ISO 16140-7 differs from the other parts in the
ISO 16140 series, as it is intended for microbial identification for which there is no reference method and,
therefore, it is not possible to run a method comparison study. The validation study in ISO 16140-7 specifies
the identification method principle, the identification database and algorithm when appropriate, and the
agar(s) from which strains can be identified. If properly characterized and successfully validated, the
identification method can only be validly used on strains recovered on the agars covered and shown to have
been acceptable within the validation study.
NOTE 3 Whole-genome sequencing (WGS) in accordance with ISO 23418 will eventually be a reference method
for all microorganisms, but the implementation of this technique is still at an early stage. Therefore, the use of WGS
cannot currently be requested as a reference method for a large panel of strains.
ISO 16140-7:2024, Figure 3, shows the possibilities where an alternative confirmation method validated
in accordance with ISO 16140-6 and an alternative identification method validated in accordance with
ISO 16140-7 can be applied within a reference method or an ISO 16140-2 validated detection or enumeration
method. The result provided by the ISO 16140-7 validated method can be considered as additional
information on the identity of the tested colony(ies); this result cannot be taken as a confirmation result.
When there is a discrepancy between the results of the ISO 16140-6 validated method and the ISO 16140-7
validated method, a root ca
...
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