ISO/FDIS 5354-1

Molecular biomarkers — Detection of DNA in cotton used for textile production — Part 1: Extraction of DNA from cottonseed and raw materials derived therefrom

This document provides requirements and recommendations to laboratories that perform PCR analyses of cottonseed, leaf, cotton fibre and cotton fibre-derived materials. The following are within the scope of this document:a) identifying the materials to be assessed, based on the probability of obtaining good quality, fit for purpose DNA from the materials in subsequent steps in the cotton cloth production process; b) specifying a method for efficient DNA isolation from cotton and cotton-derived materials; c) specifying the cotton-specific method(s) to be used as control for amplifiable DNA; d) specifying the screening procedure that provides optimal chances to detect the target DNA as a result of the performance of the lowest number of element screening assays. This document does not cover sampling of the seed, bale or fabric, or preparation of the laboratory sample. The standards sets describes the conditions for obtaining DNA for the screening of specific DNA elements and is set up in a way that offers the probability of also detecting unknown GM cotton events that possibly contain similar DNA sequences. A recommended sampling method is given in ISO 6497. General guidance for the sampling of bulk materials or for cotton-based products is available in standards such as ISO 1130, ASTM D1441-12 ISTA rules . TENTATIVE TEXTILE STANDARD No. 77. (1963)

Biomarqueurs moléculaires — Détection d’ADN dans le coton utilisé pour la production textile — Partie 1: Extraction d’ADN à partir de graines de coton et de matières premières issues de celles-ci

General Information

• Status: Not Published
• ICS: 59.080.01 - Textiles in general
• Technical Committee: ISO/TC 34/SC 16 - Horizontal methods for molecular biomarker analysis
• Drafting Committee: ISO/TC 34/SC 16 - Horizontal methods for molecular biomarker analysis
• Current Stage: 5020 - FDIS ballot initiated: 2 months. Proof sent to secretariat
• Start Date: 12-Mar-2025
• Due Date: 12-Mar-2025
• Completion Date: 12-Mar-2025

Relations

• Revises: IWA 32:2019 - Screening of genetically modified organisms (GMOs) in cotton and textiles
• Effective Date: 06-Jun-2022

Standards Content (Sample)

FINAL DRAFT
International
Standard
ISO/TC 34/SC 16
Molecular biomarkers — Detection
Secretariat: ANSI
of DNA in cotton used for textile
Voting begins on:
production —
2025-03-12
Part 1:
Voting terminates on:
2025-05-07
Extraction of DNA from cottonseed
and raw materials derived
therefrom
Biomarqueurs moléculaires — Détection d’ADN dans le coton
utilisé pour la production textile —
Partie 1: Extraction d’ADN à partir de graines de coton et de
matières premières issues de celles-ci
RECIPIENTS OF THIS DRAFT ARE INVITED TO SUBMIT,
WITH THEIR COMMENTS, NOTIFICATION OF ANY
RELEVANT PATENT RIGHTS OF WHICH THEY ARE AWARE
AND TO PROVIDE SUPPOR TING DOCUMENTATION.
IN ADDITION TO THEIR EVALUATION AS
BEING ACCEPTABLE FOR INDUSTRIAL, TECHNO-
LOGICAL, COMMERCIAL AND USER PURPOSES, DRAFT
INTERNATIONAL STANDARDS MAY ON OCCASION HAVE
TO BE CONSIDERED IN THE LIGHT OF THEIR POTENTIAL
TO BECOME STAN DARDS TO WHICH REFERENCE MAY BE
MADE IN NATIONAL REGULATIONS.
Reference number
FINAL DRAFT
International
Standard
ISO/TC 34/SC 16
Molecular biomarkers — Detection
Secretariat: ANSI
of DNA in cotton used for textile
Voting begins on:
production —
Part 1:
Voting terminates on:
Extraction of DNA from cottonseed
and raw materials derived
therefrom
Biomarqueurs moléculaires — Détection d’ADN dans le coton
utilisé pour la production textile —
Partie 1: Extraction d’ADN à partir de graines de coton et de
matières premières issues de celles-ci
RECIPIENTS OF THIS DRAFT ARE INVITED TO SUBMIT,
WITH THEIR COMMENTS, NOTIFICATION OF ANY
RELEVANT PATENT RIGHTS OF WHICH THEY ARE AWARE
AND TO PROVIDE SUPPOR TING DOCUMENTATION.
© ISO 2025
IN ADDITION TO THEIR EVALUATION AS
All rights reserved. Unless otherwise specified, or required in the context of its implementation, no part of this publication may
BEING ACCEPTABLE FOR INDUSTRIAL, TECHNO-
LOGICAL, COMMERCIAL AND USER PURPOSES, DRAFT
be reproduced or utilized otherwise in any form or by any means, electronic or mechanical, including photocopying, or posting on
INTERNATIONAL STANDARDS MAY ON OCCASION HAVE
the internet or an intranet, without prior written permission. Permission can be requested from either ISO at the address below
TO BE CONSIDERED IN THE LIGHT OF THEIR POTENTIAL
or ISO’s member body in the country of the requester.
TO BECOME STAN DARDS TO WHICH REFERENCE MAY BE
MADE IN NATIONAL REGULATIONS.
ISO copyright office
CP 401 • Ch. de Blandonnet 8
CH-1214 Vernier, Geneva
Phone: [phone]
Email: [e-mail]
Website: www.iso.org
Published in Switzerland Reference number
ii
Contents Page
Foreword .iv
Introduction .v
1 Scope . 1
2 Normative references . 1
3 Terms and definitions . 2
4 Principle . 3
5 Identification of a suitable cotton endogenous DNA marker . 3
6 Test sample preparation . 3
7 Assessment of DNA extraction methods for different cotton production stages . 4
7.1 General .4
7.2 Results from the single laboratory analysis of DNA extraction methods .4
7.3 Conclusion .5
8 Storage . 5
9 DNA quantitation . 5
10 DNA quality control. 5
10.1 General .5
10.2 Use of SAH7 marker as a cotton DNA quality control assay .5
10.3 Analysis for PCR inhibitors.6
10.4 Cotton matrix control method . .6
10.4.1 Results .6
11 Test report . 6
Annex A (informative) Cotton endogenous control analysis . 8
Annex B (informative) Assessment of DNA extraction methods for different cotton production
stages . 10
Annex C (informative) PCR method to detect SAH 7 gene target DNA in cotton . 14
Annex D (informative) Evaluation of DNA isolated with a commercial spin column-based DNA
extraction system designed for the extraction of DNA from stool samples with the SAH 7
method . 16
Bibliography .20

iii
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 16,
Horizontal methods for molecular biomarker analysis.
This first edition, along with ISO/TS 5354-2:2024, cancels and replaces IWA 32:2019, which has been
technically revised throughout.
A list of all parts in the ISO 5354 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.

iv
Introduction
The purpose of this document is to provide guidance to assess whether cotton, cotton fibre or cotton-derived
materials, or all of these, contain a specific DNA sequence or sequences. This guidance can be applied to
detection of pure genetically modified (GM) cotton in textile production, detection of a specific GM cotton
target sequence in other cotton and for confirming or tracing a particular species, variety or genetic marker.
[1]
While GM-cotton cultivation covers a large percentage of global cotton production today, there are
countries where the cultivation of GM cotton is not permitted by law, as well as voluntary, private and public
standards that do not permit the intentional use of genetically modified organisms (GMOs) in the cotton
and textile production process or require labelling. Due to asynchronous regulatory approvals, a GM cotton
variety that is approved for growth and import and in one country can be disapproved or require labelling
in another country. There has been a need for detection of a specific GM cotton event in GM (or non-GM)
cotton. The detection methods submitted and approved by global regulatory agencies are available for that
purpose and this method does not supplant those nor the results of those analyses.
Growers of non-GM cotton can provide traceability and certification of cottonseed to ensure that the seeds
entering a certified cultivation scheme are not GM. If the starter seed is conventional (non-GM), which can
be accurately determined depending on the availability of methods, and growers follow their certification
process then the ginned fibre can be received as is without misleading consumers. This document provides
evidence that DNA extraction methods are only effective and accurate for seeds and leaves. Although pure
GM cotton can be detected at the ginned cotton stage, and potentially at the griege yarn stage, non-GM
cotton cannot be claimed if a negative result is obtained because there is a significant potential for a false
negative result due to the lack of polymerase chain reaction (PCR) quality DNA.
The DNA sequence screening approach described in this document is based on PCR-methods. The methods
described in this document are designed to work on all four of the major commercial cotton species:
Gossypium hirsutum, G. barbadense, G. arboreum and G. herbaceum.
Cotton (Gossypium spp.) has been cultivated for lint for over 8 000 years. There are over 50 species in the
[2] [3]
Gossypium genus. The Gossypium genome is complex, containing 2,25 to 2,43 gigabasepairs.
This document describes the key factors necessary to screen cottonseed, cotton leaf and fibre samples at
different stages of textile development in the cotton production chain for the potential presence of specific
DNA elements. The protocol describes two major steps:
a) an effective way to isolate DNA from cotton materials;
b) a method to confirm that the isolated DNA is PCR quality DNA, i.e. suitable for PCR (preferred markers
chosen for this purpose will be nuclear, and low copy number).
[4]
GM element screening is described in ISO/TS 5354-2 .
The single laboratory validation studies described in this document including method development was
carried out by the Wageningen University and Research Institute (RIKILT), the Netherlands.

v
...

ISO/DIS FDIS 5354-1.2:2025(E)
Date: 2025-01-21
ISO/TC 34/SC 16/JWG 12
Secretariat: ANSI
Date: 2025-02-25
Molecular biomarkers — Detection of DNA in cotton used for textile
production —
Part 1:
Extraction of DNA from cottonseed and raw materials derived
therefrom
Biomarqueurs moléculaires — Détection d’ADN dans le coton utilisé pour la production textile —
Partie 1: Extraction d’ADN à partir de graines de coton et de matières premières issues de celles-ci
FDIS stage
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
E-mail: [e-mail]
Website: www.iso.org
Published in Switzerland
ii
Contents
Foreword . iv
Introduction . v
1 Scope . 1
2 Normative references . 1
3 Terms and definitions . 2
4 Principle . 3
5 Identification of a suitable cotton endogenous DNA marker . 3
6 Test sample preparation . 4
7 Assessment of DNA extraction methods for different cotton production stages . 4
7.1 General . 4
7.2 Results from the single laboratory analysis of DNA extraction methods . 5
7.3 Conclusion . 5
8 Storage . 6
9 DNA quantitation . 6
10 DNA quality control . 6
10.1 General . 6
10.2 Use of SAH7 marker as a cotton DNA quality control assay . 6
10.3 Analysis for PCR inhibitors . 6
10.4 Cotton matrix control method . 6
11 Test report . 7
Annex A (informative) Cotton endogenous control analysis . 8
Annex B (informative) Assessment of DNA extraction methods for different cotton production
stages . 10
Annex C (informative) PCR method to detect SAH 7 gene target DNA in cotton . 14
Annex D (informative) Evaluation of DNA isolated with a commercial spin column-based DNA
extraction system designed for the extraction of DNA from stool samples with the SAH 7
method . 17
Bibliography . 22

iii
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 documentsdocument 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 16,
Horizontal methods for molecular biomarker analysis.
This first edition, along with ISO/TS 5354-2:2024, cancels and replaces IWA 32:2019, which has been
technically revised throughout.
A list of all parts in the ISO 5354 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.
iv
Introduction
The purpose of this document is to provide guidance to assess whether cotton, cotton fibre or cotton-derived
materials, or all of these, contain a specific DNA sequence or sequences. This guidance can be applied to
detection of pure genetically modified (GM) cotton in textile production, detection of a specific GM cotton
target sequence in other cotton and for confirming or tracing a particular species, variety or genetic marker.
[1]
While GM-cotton cultivation covers a large percentage of global cotton production today, there are countries
where the cultivation of GM cotton is not permitted by law, as well as voluntary, private and public standards
that do not permit the intentional use of genetically modified organisms (GMOs) in the cotton and textile
production process or require labelling. Due to asynchronous regulatory approvals, a GM cotton variety that
is approved for growth and import and in one country can be disapproved or require labelling in another
country. There has been a need for detection of a specific GM cotton event in GM (or non-GM) cotton. The
detection methods submitted and approved by global regulatory agencies are available for that purpose and
this method does not supplant those nor the results of those analyses.
Growers of non-GM cotton can provide traceability and certification of cottonseed to ensure that the seeds
entering ana certified cultivation scheme are not GM. If the starter seed is conventional (non-GM), which can
be accurately determined depending on the availability of methods, and growers follow their certification
process then the ginned fibre can be received as is without misleading consumers. This document provides
evidence that DNA extraction methods are only effective and accurate for seeds and leaves. Although, pure GM
cotton can be detected at the ginned cotton stage, and potentially at the griege yarn stage, non-GM cotton
cannot be claimed if a negative result is obtained because there is a significant potential offor a false negative
result due to the lack of polymerase chain reaction (PCR) quality DNA.
The DNA sequence screening approach described in this document is based on PCR-methods. The methods
described in this document are designed to work on all four of the major commercial cotton species: Gossypium
hirsutum, G. barbadense, G. arboreum and G. herbaceum.
Cotton (Gossypium spp.) has been cultivated for lint for over 8 000 years. There are over 50 species in the
[2] [3]
Gossypium genus. The Gossypium genome is complex, containing 2,25 to 2,43 gigabasepairs.
This document describes the key factors necessary to screen cottonseed, cotton leaf and fibre samples at
different stages of textile development in the cotton production chain for the potential presence of specific
DNA elements. The protocol describes two major steps:
a) an effective way to isolate DNA from cotton materials;
b) a method to confirm that the isolated DNA is PCR quality DNA, i.e. suitable for PCR (preferred markers
chosen for this purpose will be nuclear, and low copy number).
[4]
Genetically modifiedGM element screening is described in ISO/TS 5354-2 .
The single laboratory validation studies described in this document including method development was
carried out by the Wageningen University and Research Institute (RIKILT), the Netherlands.
v
Molecular biomarkers — Detection of DNA in cotton used for textile
production —
Part 1:
Extraction of DNA from cottonseed and raw materials derived
therefrom
1 Scope
This document specifies requirements and recommendations to laboratories that perform extraction of
PCRpolymerase chain reaction (PCR) quality deoxyribonucleic acid (DNA) from cottonseed, cotton leaf and
raw material derived therefrom, that is sufficient for the purpose of polymerase chain reaction (PCR) analysis.
This document is applicable to:
a) identifying cotton raw material from which PCR quality DNA can be extracted;
b) specifying a method for effective DNA extraction from cotton and cotton-derived raw materials;
c) specifying the cotton-specific marker(s) to be used as controls for PCR amplification of DNA.
A PCR result obtained from analysis of cottonseed, cotton leaf and to some extant raw materials derived
therefrom can only indicate that it is not derived from pure genetically modified organism (GMO) - )-derived
cotton. Admixtures of GMO-derived cotton cannot be detected for cotton fibre and cotton fibre-derived
materials.
This document does not apply to bulk sampling of the seed, bale or processed fabric and yarn. A recommended
[5]
sampling method is given in ISO 6497 . General guidance for the sampling of bulk materials or for cotton-
[6] [7]
based products is available in standards such as ASTM D1441-12 and CEN/TS 15568 .
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 3696:1987, Water for analytical laboratory use — Specification and test methods
ISO 16577, Molecular biomarker analysis — Vocabulary for molecular biomarker analytical methods in
agriculture and food production
ISO 21570:2005, Foodstuffs — Methods of analysis for the detection of genetically modified organisms and
derived products — Quantitative nucleic acid based methods
ISO 21571, Foodstuffs — Methods of analysis for the detection of genetically modified organisms and derived
products — Nucleic acid extraction
ISO 24276:2006, Foodstuffs — Methods of analysis for the detection of genetically modified organisms and
derived products — General requirements and definitions
ISO 24276:2006/Amd 1:2013, Foodstuffs — Methods of analysis for the detection of genetically modified
organisms and derived products — General requirements and definitions — Amendment 1
3 Terms and definitions
For the purposes of this document, the terms and definitions given in ISO 16577 Molecular biomarker analysis
— Vocabulary for molecular biomarker analytical methods in agriculture and food production and the
following apply.
ISO and IEC maintain terminology databases for use in standardization at the following addresses:
— ISO Online
...

PROJET FINAL
Norme
internationale
ISO/TC 34/SC 16
Biomarqueurs moléculaires —
Secrétariat: ANSI
Détection d’ADN dans le coton
Début de vote:
utilisé pour la production textile —
2025-03-12
Partie 1:
Vote clos le:
2025-05-07
Extraction d’ADN à partir de graines
de coton et de matières premières
issues de celles-ci
Molecular biomarkers — Detection of DNA in cotton used for
textile production —
Part 1: Extraction of DNA from cottonseed and raw materials
derived therefrom
LES DESTINATAIRES DU PRÉSENT PROJET SONT
INVITÉS À PRÉSENTER, AVEC LEURS OBSERVATIONS,
NOTIFICATION DES DROITS DE PROPRIÉTÉ DONT ILS
AURAIENT ÉVENTUELLEMENT CONNAISSANCE ET À
FOURNIR UNE DOCUMENTATION EXPLICATIVE.
OUTRE LE FAIT D’ÊTRE EXAMINÉS POUR
ÉTABLIR S’ILS SONT ACCEPTABLES À DES FINS
INDUSTRIELLES, TECHNOLOGIQUES ET COM-MERCIALES,
AINSI QUE DU POINT DE VUE DES UTILISATEURS, LES
PROJETS DE NORMES
INTERNATIONALES DOIVENT PARFOIS ÊTRE CONSIDÉRÉS
DU POINT DE VUE DE LEUR POSSI BILITÉ DE DEVENIR DES
NORMES POUVANT
SERVIR DE RÉFÉRENCE DANS LA RÉGLEMENTATION
NATIONALE.
Numéro de référence
PROJET FINAL
Norme
internationale
ISO/TC 34/SC 16
Biomarqueurs moléculaires —
Secrétariat: ANSI
Détection d’ADN dans le coton
Début de vote:
utilisé pour la production textile —
2025-03-12
Partie 1:
Vote clos le:
2025-05-07
Extraction d’ADN à partir de graines
de coton et de matières premières
issues de celles-ci
Molecular biomarkers — Detection of DNA in cotton used for
textile production —
Part 1: Extraction of DNA from cottonseed and raw materials
derived therefrom
LES DESTINATAIRES DU PRÉSENT PROJET SONT
INVITÉS À PRÉSENTER, AVEC LEURS OBSERVATIONS,
NOTIFICATION DES DROITS DE PROPRIÉTÉ DONT ILS
AURAIENT ÉVENTUELLEMENT CONNAISSANCE ET À
FOURNIR UNE DOCUMENTATION EXPLICATIVE.
DOCUMENT PROTÉGÉ PAR COPYRIGHT
OUTRE LE FAIT D’ÊTRE EXAMINÉS POUR
ÉTABLIR S’ILS SONT ACCEPTABLES À DES FINS
© ISO 2025 INDUSTRIELLES, TECHNOLOGIQUES ET COM-MERCIALES,
AINSI QUE DU POINT DE VUE DES UTILISATEURS, LES
Tous droits réservés. Sauf prescription différente ou nécessité dans le contexte de sa mise en œuvre, aucune partie de cette
PROJETS DE NORMES
INTERNATIONALES DOIVENT PARFOIS ÊTRE CONSIDÉRÉS
publication ne peut être reproduite ni utilisée sous quelque forme que ce soit et par aucun procédé, électronique ou mécanique,
DU POINT DE VUE DE LEUR POSSI BILITÉ DE DEVENIR DES
y compris la photocopie, ou la diffusion sur l’internet ou sur un intranet, sans autorisation écrite préalable. Une autorisation peut
NORMES POUVANT
être demandée à l’ISO à l’adresse ci-après ou au comité membre de l’ISO dans le pays du demandeur.
SERVIR DE RÉFÉRENCE DANS LA RÉGLEMENTATION
NATIONALE.
ISO copyright office
Case postale 401 • Ch. de Blandonnet 8
CH-1214 Vernier, Genève
Tél.: [phone]
E-mail: [e-mail]
Web: www.iso.org
Publié en Suisse Numéro de référence
ii
Sommaire Page
Avant-propos .iv
Introduction .v
1 Domaine d’application . 1
2 Références normatives . 1
3 Termes et définitions . 2
4 Principe. 3
5 Identification d’un marqueur d’ADN endogène approprié du coton . 3
6 Préparation de l’échantillon pour essai . 4
7 Évaluation des méthodes d’extraction d’ADN pour différentes phases de production du
coton . 4
7.1 Généralités .4
7.2 Résultats de l’analyse intralaboratoire des méthodes d’extraction d’ADN .5
7.3 Conclusion .5
8 Stockage . 5
9 Quantification de l’ADN . 5
10 Contrôle de la qualité de l’ADN . 6
10.1 Généralités .6
10.2 Utilisation du marqueur SAH7 comme essai de contrôle de la qualité de l’ADN du coton . .6
10.3 Analyse des inhibiteurs de PCR .6
10.4 Méthode de contrôle de matrice de coton .6
10.4.1 Résultats .6
11 Rapport d’essai . 7
Annexe A (informative) Analyse des témoins endogènes du coton . 8
Annexe B (informative) Évaluation des méthodes d’extraction d’ADNpour différentes phases
de production du coton . 10
Annexe C (informative) Méthode de PCR pour détecter l’ADN cible du gène SAH 7 dans le coton . 14
Annexe D (informative) Évaluation de l’ADN extrait à l’aide d’un système commercial
d’extraction d’ADN sur colonne à centrifuger conçu pour l’extraction d’ADNà partir
d’échantillons de selles avec la méthode SAH 7 . 17
Bibliographie .22

iii
Avant-propos
L’ISO (Organisation internationale de normalisation) est une fédération mondiale d’organismes nationaux
de normalisation (comités membres de l’ISO). L’élaboration des Normes internationales est en général
confiée aux comités techniques de l’ISO. Chaque comité membre intéressé par une étude a le droit de faire
partie du comité technique créé à cet effet. Les organisations internationales, gouvernementales et non
gouvernementales, en liaison avec l’ISO participent également aux travaux. L’ISO collabore étroitement avec
la Commission électrotechnique internationale (IEC) en ce qui concerne la normalisation électrotechnique.
Les procédures utilisées pour élaborer le présent document et celles destinées à sa mise à jour sont
décrites dans les Directives ISO/IEC, Partie 1. Il convient, en particulier, de prendre note des différents
critères d’approbation requis pour les différents types de documents ISO. Le présent document
a été rédigé conformément aux règles de rédaction données dans les Directives ISO/IEC, Partie 2
(voir www.iso.org/directives).
L’ISO attire l’attention sur le fait que la mise en application du présent document peut entraîner l’utilisation
d’un ou de plusieurs brevets. L’ISO ne prend pas position quant à la preuve, à la validité et à l’applicabilité de
tout droit de brevet revendiqué à cet égard. À la date de publication du présent document, l’ISO n’avait pas
reçu notification qu’un ou plusieurs brevets pouvaient être nécessaires à sa mise en application. Toutefois,
il y a lieu d’avertir les responsables de la mise en application du présent document que des informations
plus récentes sont susceptibles de figurer dans la base de données de brevets, disponible à l’adresse
www.iso.org/brevets. L’ISO ne saurait être tenue pour responsable de ne pas avoir identifié de tels droits
de propriété.
Les appellations commerciales éventuellement mentionnées dans le présent document sont données pour
information, par souci de commodité, à l’intention des utilisateurs et ne sauraient constituer un engagement.
Pour une explication de la nature volontaire des normes, la signification des termes et expressions
spécifiques de l’ISO liés à l’évaluation de la conformité, ou pour toute information au sujet de l’adhésion de
l’ISO aux principes de l’Organisation mondiale du commerce (OMC) concernant les obstacles techniques au
commerce (OTC), voir www.iso.org/avant-propos.
Le présent document a été élaboré par le comité technique ISO/TC 34, Produits alimentaires, sous-
comité SC 16, Méthodes horizontales pour l’analyse de biomarqueurs moléculaires.
Cette première édition, conjointement avec l’ISO/TS 5354-2:2024, annule et remplace l’IWA 32:2019, qui a
fait l’objet d’une révision technique complète.
Une liste de toutes les parties de la série ISO 5354 se trouve sur le site web de l’ISO.
Il convient que l’utilisateur adresse tout retour d’information ou toute question concernant le présent
document à l’organisme national de normalisation de son pays. Une liste exhaustive desdits organismes
se trouve à l’adresse www.iso.org/fr/members.html.

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Introduction
Le présent document vise à fournir des recommandations pour évaluer si du coton, des fibres de coton et/
ou des matériaux dérivés du coton contiennent une ou plusieurs séquences d’ADN spécifiques. Ce document
d’orientation peut être appliqué à la détection de coton génétiquement modifié (GM) pur dans la production
textile, à la détection d’une séquence cible particulière de coton dans un autre type de coton GM et pour
confirmer ou tracer une espèce ou une variété donnée ou un marqueur génétique particulier.
À l’heure actuelle, la culture du coton GM représente un pourcentage important de la production mondiale
[1]
de coton . Toutefois, dans certains pays, la culture du coton GM est interdite par la loi ainsi que par des
normes publiques et privées de nature volontaire qui interdisent l’usage intentionnel d’organismes
génétiquement modifiés (OGM) dans le cadre du processus de production de coton et de textile ou exigent
un étiquetage en conséquence. Du fait de la nature asynchrone des autorisations réglementaires, une variété
de coton GM dont la culture et l’importation sont autorisées dans un pays peut ne pas être autorisée ou
devoir être étiquetée en conséquence dans un autre pays. Il a été nécessaire de détecter un événement de
coton GM spécifique dans le coton GM (ou non GM). Les méthodes de détection soumises et approuvées par
les organismes de réglementation internationaux sont disponibles à cette fin. Cette méthode ne remplace
ni ces méthodes, ni les résultats de ces analyses.
Les producteurs de coton non GM peuvent fournir une traçabilité et une certification des graines de coton
pour garantir que les graines entrant dans un principe de culture certifiée ne sont pas GM. Si la graine
de départ est conventionnelle (non GM), ce qui peut être déterminé avec exactitude selon la disponibilité
de méthodes, et si les producteurs suivent leur processus de certification, la fibre égrenée peut alors être
certifiée non GM sans induire les consommateurs en erreur. Le présent document démontre que les méthodes
d’extraction d’ADN ne sont efficaces et exactes que pour les graines et les feuilles. Même s’il est possible
de détecter le coton GM pur au stade de coton égrené et potentiellement au stade de fil grège, un coton
pour lequel un résultat négatif est obtenu ne peut pas être déclaré non GM en raison du risque significatif
de résultat faussement négatif dû à une insuffisance d’ADN de qualité PCR (réaction de polymérisation en
chaîne).
L’approche de criblage de séquence d’ADN décrite dans le présent document repose sur des méthodes PCR.
Les méthodes décrites dans le présent document sont conçues pour être applicables aux quatre principales
espèces de coton du commerce: Gossypium hirsutum, G. barbadense, G. arbor
...