SIST EN ISO 11781:2025

SLOVENSKI STANDARD
01-junij-2025
Nadomešča:
SIST-TS CEN/TS 17329-1:2021
Analiza molekularnih biomarkerjev - Zahteve in navodila za validacijo kvalitativnih
metod s polimerazno verižno reakcijo (PCR) v realnem času v posameznem
laboratoriju (ISO 11781:2025)
Molecular biomarker analysis - Requirements and guidance for single-laboratory
validation of qualitative real-time polymerase chain reaction (PCR) methods (ISO
11781:2025)
Untersuchung auf molekulare Biomarker - Anforderungen und Leitfaden für die
Einzellaborvalidierung qualitativer Real-time-PCR-Verfahren (ISO 11781:2025)
Analyse de biomarqueurs moléculaires - Exigences et recommandations pour la
validation intralaboratoire des méthodes de PCR qualitative en temps réel (ISO
11781:2025)
Ta slovenski standard je istoveten z: EN ISO 11781:2025
ICS:
67.050 Splošne preskusne in General methods of tests and
analizne metode za živilske analysis for food products
proizvode
2003-01.Slovenski inštitut za standardizacijo. Razmnoževanje celote ali delov tega standarda ni dovoljeno.

EN ISO 11781
EUROPEAN STANDARD
NORME EUROPÉENNE
April 2025
EUROPÄISCHE NORM
ICS 67.050 Supersedes CEN/TS 17329-1:2021
English Version
Molecular biomarker analysis - Requirements and
guidance for single-laboratory validation of qualitative
real-time polymerase chain reaction (PCR) methods (ISO
11781:2025)
Analyse de biomarqueurs moléculaires - Exigences et Untersuchung auf molekulare Biomarker -
recommandations pour la validation intralaboratoire Anforderungen und Leitfaden für die
des méthodes de PCR qualitative en temps réel (ISO Einzellaborvalidierung qualitativer Real-time-PCR-
11781:2025) Verfahren (ISO 11781:2025)
This European Standard was approved by CEN on 4 April 2025.

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
© 2025 CEN All rights of exploitation in any form and by any means reserved Ref. No. EN ISO 11781:2025 E
worldwide for CEN national Members.

Contents Page
European foreword . 3

European foreword
This document (EN ISO 11781:2025) has been prepared by Technical 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 endorsement, at the latest by October 2025, and conflicting national standards shall
be withdrawn at the latest by October 2025.
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 CEN/TS 17329-1:2021.
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 11781:2025 has been approved by CEN as EN ISO 11781:2025 without any modification.

International
Standard
ISO 11781
First edition
Molecular biomarker analysis —
2025-04
Requirements and guidance for
single-laboratory validation of
qualitative real-time polymerase
chain reaction (PCR) methods
Analyse de biomarqueurs moléculaires — Exigences et
recommandations pour la validation intralaboratoire des
méthodes de PCR qualitative en temps réel
Reference number
ISO 11781:2025(en) © ISO 2025
ISO 11781:2025(en)
© ISO 2025
All rights reserved. Unless otherwise specified, or required in the context of its implementation, no part of this publication may
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Phone: +41 22 749 01 11
Email: copyright@iso.org
Website: www.iso.org
Published in Switzerland
ii
ISO 11781:2025(en)
Contents Page
Foreword .iv
Introduction .v
1 Scope . 1
2 Normative references . 1
3 Terms and definitions . 1
4 Principle . 2
5 Single-laboratory validation of the performance characteristics . 2
5.1 General .2
5.2 Limit of detection .2
5.3 Determining copies of DNA target sequences in DNA test materials .3
5.4 Evaluation of data for the limit of detection .3
5.5 PCR efficiency and variability of the measured copy number around the limit of
detection .4
5.6 Specificity .4
5.6.1 General .4
5.6.2 Bioinformatic (in silico) test for specificity .4
5.6.3 Practical test for specificity .4
5.6.4 Robustness . .5
6 Validation report . 6
Annex A (informative) Estimation of the number of copies of the DNA target sequence . 7
Annex B (informative) Determination of limit of detection, precision and PCR efficiency . 9
Annex C (informative) Generalized linear mixed model with log-log link .13
Annex D (informative) Robustness testing . 19
Bibliography .21

iii
ISO 11781:2025(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,
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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.
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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 16,
Horizontal methods for molecular biomarker analysis, in collaboration with the European Committee for
Standardization (CEN) Technical Committee CEN/TC 275, Food analysis - Horizontal methods, in accordance
with the Agreement on technical cooperation between ISO and CEN (Vienna Agreement).
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
ISO 11781:2025(en)
Introduction
Qualitative real-time polymerase chain reaction (PCR) methods currently find broad application for the
detection of specific DNA sequences in food (e.g. for the detection and identification of genetically modified
organisms and the products derived thereof, for food authentication and speciation and for other purposes). It
is important that a newly developed food analytical method is fit for purpose and meets certain performan
...