ISO 6579-4

Microbiology of the food chain — Horizontal method for the detection, enumeration and serotyping of Salmonella — Part 4: Identification of monophasic Salmonella Typhimurium (1,4,[5],12:i:-) by polymerase chain reaction (PCR)

Microbiologie de la chaîne alimentaire — Méthode horizontale pour la recherche, le dénombrement et le sérotypage des Salmonella — Partie 4: Identification du variant monophasique de Salmonella Typhimurium (1,4,[5],12:i:-) par réaction de polymérisation en chaîne (PCR)

General Information

• Status: Not Published
• ICS: 07.100.30 - Food microbiology
• Technical Committee: ISO/TC 34/SC 9 - Microbiology
• Drafting Committee: ISO/TC 34/SC 9 - Microbiology
• Current Stage: 6000 - International Standard under publication
• Due Date: 27-Dec-2024
• Completion Date: 27-Dec-2024

Relations

• Consolidates: ISO 3631:2019 - Citrus fruits — Guidelines for storage
• Effective Date: 06-Jun-2022

Standards Content (Sample)

FINAL DRAFT
International
Standard
ISO/FDIS 6579-4
ISO/TC 34/SC 9
Microbiology of the food chain —
Secretariat: AFNOR
Horizontal method for the
Voting begins on:
detection, enumeration and
2024-10-31
serotyping of Salmonella —
Voting terminates on:
2024-12-26
Part 4:
Identification of monophasic
Salmonella Typhimurium
(1,4,[5],12:i:-) by polymerase
chain reaction (PCR)
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-
ISO/CEN PARALLEL PROCESSING 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
ISO/FDIS 6579-4:2024(en) © ISO 2024

ISO/FDIS 6579-4:2024(en)
© ISO 2024
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: [phone]
Email: [e-mail]
Website: www.iso.org
Published in Switzerland
ii
ISO/FDIS 6579-4:2024(en)
Contents Page
Foreword .iv
Introduction .v
1 Scope . 1
2 Normative references . 1
3 Terms and definitions . 2
4 Principle . 2
4.1 General .2
4.2 Preparation of well-isolated colonies .2
4.3 Suspension of a colony .2
4.4 Amplification and detection .2
5 Culture media and reagents . 3
6 Equipment and consumables . 3
7 Presumptive monophasic Salmonella Typhimurium . 3
8 Culturing the isolate . 4
9 Procedure . 4
9.1 Preparation of cell suspension or DNA .4
9.2 PCR amplification and detection .4
9.2.1 General .4
9.2.2 PCR controls .4
10 Expression of results . 4
11 Performance characteristics of the method . 5
11.1 Validation in accordance with ISO 17468 .5
11.2 Performance characteristics .5
11.2.1 Method(s) evaluation study .5
11.2.2 Interlaboratory study .5
12 Test report . 6
13 Quality assurance . 6
Annex A (normative) Culture media and reagents. 7
Annex B (informative) Probe-based multiplex real-time PCR assay for the identification of
monophasic Salmonella Typhimurium (1,4,[5],12:i:-) . 9
Annex C (informative) Agarose gel-based multiplex target PCR assay for the identification of
monophasic Salmonella Typhimurium (1,4,[5],12:i:-) .13
Annex D (informative) Agarose gel-based single target PCR assay for the identification of
monophasic Salmonella Typhimurium (1,4,[5],12:i:-) .18
Annex E (informative) Performance characteristics .25
Bibliography .32

iii
ISO/FDIS 6579-4:2024(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 6579 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
ISO/FDIS 6579-4:2024(en)
Introduction
In several international, regional and national laws, regulatory limits are set to ensure the so-called
“absence” of Salmonella spp. in samples of the food chain. Moreover, several European Commission (EC)
regulations also demand the absence of particular Salmonella serovars which have shown to cause a
relatively high percentage of human salmonellosis. One of these Salmonella serovars for which legal criteria
are set is Salmonella Typhimurium, including its monophasic variant 1,4,[5],12:i:- (e.g. Regulation (EC)
[10]
No. 1086/2011 ). Hence, it is important to know that a serovar found with antigenic formula 1,4,[5],12:i:-
is indeed the monophasic variant of Salmonella Typhimurium (1,4,[5],12:i:1,2) and not the monophasic
variant of another Salmonella (S.) serovar for which no criteria are set, such as S. Lagos (1,4,[5],12:i:1,5),
S. Agama (4,12:i:1,6), S. Farsta (4,[5],12:i:e,n,x), S. Tsevie (1,4,12:i:e,n,z ), S. Gloucester (1,4,12,27:i:l,w) or
S. Tumodi (1,4,12:i:z ). Confirmational distinction between Salmonella Typhimurium and Salmonella non-
Typhimurium serovars can be determined using molecular analysis, such as the PCR technique(s) described
in this document.
v
FINAL DRAFT International Standard ISO/FDIS 6579-4:2024(en)
Microbiology of the food chain — Horizontal method for the
detection, enumeration and serotyping of Salmonella —
Part 4:
Identification of monophasic Salmonella Typhimurium
(1,4,[5],12:i:-) by polymerase chain reaction (PCR)
WARNING — In order to safeguard the health of laboratory personnel, it is essential that tests for
detecting, enumerating and (sero)typing Salmonella are only undertaken in properly equipped
laboratories, under the control of a skilled microbiologist, and that great care is taken in the disposal
of all incubated materials. Persons using this document should be familiar with normal laboratory
practice. This document does not purport to address all of the safety aspects, if any, associated with
its use. It is the responsibility of the user to establish appropriate safety and health practices.
1 Scope
This document specifies a horizontal in vitro method for the molecular identification and differentiation of
the monophasic variant of Salmonella enterica subsp. enterica serovar Typhimurium (1,4,[5],12:i:-) lacking
the second H phase H:1,2, starting from isolates. The method detects specific DNA sequences of an intergenic
region of the first H phase flagellin cluster for identification of Salmonella enterica subsp. enterica serovar
Typhimurium (further called Salmonella Typhimurium) and specific DNA sequences of genes associated
with second H phase flagellar antigen expression.
The method is applicable for:
— differentiation of the isolate under analysis between monophasic Salmonella Typhimurium and the
monophasic variant of another Salmonella non-Typhimurium serovar that has the same antigenic formula;
— identification of the isolate under analysis being either monophasic Salmonella Typhimurium o r (biphasi
...

ISO/FDIS 6579-4:2024(E)
Date: 2024-09-10
ISO/TC 34/SC 9/WG 10
Secretariat: AFNOR
Date: 2024-10-17
Microbiology of the food chain — Horizontal method for the
detection, enumeration and serotyping of Salmonella —
Part 4:
Identification of monophasic Salmonella Typhimurium (1,4,[5],12:i:-)
by polymerase chain reaction (PCR)
FDIS stage
ISO/FDIS 6579-4:2024(en)
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
ISO/FDIS 6579-4:2024(en)
Contents
Foreword . iv
Introduction . v
1 Scope . 1
2 Normative references . 1
3 Terms and definitions . 2
4 Principle . 2
4.1 General . 2
4.2 Preparation of well-isolated colonies . 2
4.3 Suspension of a colony . 2
4.4 Amplification and detection . 3
5 Culture media and reagents . 3
6 Equipment and consumables. 3
7 Presumptive monophasic Salmonella Typhimurium . 4
8 Culturing the isolate. 4
9 Procedure . 4
9.1 Preparation of cell suspension or DNA . 4
9.2 PCR amplification and detection . 4
10 Expression of results . 5
11 Performance characteristics of the method . 5
11.1 Validation in accordance with ISO 17468 . 5
11.2 Performance characteristics . 5
12 Test report . 6
13 Quality assurance . 6
Annex A (normative) Culture media and reagents . 7
Annex B (informative) Probe-based multiplex real-time PCR assay for the identification of
monophasic Salmonella Typhimurium (1,4,[5],12:i:-) . 9
Annex C (informative) Agarose gel-based multiplex target PCR assay for the identification of
monophasic Salmonella Typhimurium (1,4,[5],12:i:-) . 14
Annex D (informative) Agarose gel-based single target PCR assay for the identification of
monophasic Salmonella Typhimurium (1,4,[5],12:i:-) . 20
Annex E (informative) Performance characteristics . 28
Bibliography . 35

iii
ISO/FDIS 6579-4:2024(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 6579 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
ISO/FDIS 6579-4:2024(en)
Introduction
In several international, regional and national laws, regulatory limits are set to ensure the so-called “absence”
of Salmonella spp. in samples of the food chain. Moreover, several European Commission (EC) regulations also
demand the absence of particular Salmonella serovars which have shown to cause a relatively high percentage
of human salmonellosis. One of these Salmonella serovars for which legal criteria are set is Salmonella
[10][11]
Typhimurium, including its monophasic variant 1,4,[5],12:i:- (e.g. Regulation (EC) No. 1086/2011 ).
Hence, it is important to know that a serovar found with antigenic formula 1,4,[5],12:i:- is indeed the
monophasic variant of Salmonella Typhimurium (1,4,[5],12:i:1,2) and not the monophasic variant of another
Salmonella (S.) serovar for which no criteria are set, such as S. Lagos (1,4,[5],12:i:1,5), S. Agama (4,12:i:1,6), S.
Farsta (4,[5],12:i:e,n,x), S. Tsevie (1,4,12:i:e,n,z ), S. Gloucester (1,4,12,27:i:l,w) or S. Tumodi (1,4,12:i:z ).
15 6
Confirmational distinction between Salmonella Typhimurium and Salmonella non-Typhimurium serovars can
be determined using molecular analysis, such as the PCR technique(s) described in this document.
v
Microbiology of the food chain — Horizontal method for the detection,
enumeration and serotyping of Salmonella —
Part 4:
Identification of monophasic Salmonella Typhimurium (1,4,[5],12:i:-)
by polymerase chain reaction (PCR)
WARNING — In order to safeguard the health of laboratory personnel, it is essential that tests for
detecting, enumerating and (sero)typing Salmonella are only undertaken in properly equipped
laboratories, under the control of a skilled microbiologist, and that great care is taken in the disposal
of all incubated materials. Persons using this document should be familiar with normal laboratory
practice. This document does not purport to address all of the safety aspects, if any, associated with its
use. It is the responsibility of the user to establish appropriate safety and health practices.
1 Scope
This document specifies a horizontal in vitro method for the molecular identification and differentiation of the
monophasic variant of Salmonella enterica subsp. enterica serovar Typhimurium (1,4,[5],12:i:-) lacking the
second H phase H:1,2, starting from isolates. The method detects specific DNA sequences of an intergenic
region of the first H phase flagellin cluster for identification of Salmonella enterica subsp. enterica serovar
Typhimurium (further called Salmonella Typhimurium) and specific DNA sequences of genes associated with
second H phase flagellar antigen expression.
The method is applicable for:
— differentiation of the isolate under analysis between monophasic Salmonella Typhimurium and the
monophasic variant of another Salmonella non-Typhimurium serovar that has the same antigenic formula;
— identification of the isolate under analysis being either monophasic Salmonella Typhimurium or
(biphasic) Salmonella Typhimurium.
This document is applicable for the analysis of a pure culture belonging to the genus Salmonella, isolated from:
— products intended for human consumption;
— products intended for animal feeding;
— environmental samples in the area of food and feed production and handling;
— samples from the primary production stage.
This document can also be applied in other domains for identification of monophasic Salmonella Typhimurium
(e.g. environmental, human health, animal health).
NOTE This method has been validated in a method evaluation study and in an interlaboratory study with a large set
of different strains (target and non-target strains), isolated from different sources (food products, animals, animal feed,
primary production samples and humans). For detailed information on the validation, see Annex E.
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/FDIS 6579-4:2024(en)
ISO 7218, Microbiology of the food chain — General requirements and guidance for microbiological
examinations
ISO 11133, Microbiology of food, animal feed and water — Preparation, production, storage and performance
testing of culture media
ISO 20836, Microbiology of the food chain — Polymerase chain reaction (PCR) for the detection of
microorganisms — Thermal performance testing of thermal cyclers
ISO 22174, Microbiology of the food chain — Polymerase chain reaction (PCR) for the detection and
quantification of microorganisms — General requirements and definitions
3 Terms and definitions
For the purposes of this document, the terms and definitions given in ISO 22174 and the following apply.
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
monophasic Salmonella Typhimurium
variant of Salmonella enterica subsp. enterica serovar Typhimurium lacking the second H phase H:1,2, having
the antigenic formula 1,4,[5],12:i:-
3.2
presumptive monophasic Salmonella Typhimurium
pure culture characterized as belonging to the genus Salmonella, giving a positive reaction for O-antigen O:4
and H-antigen H:i and with a negative reaction for the second H phase H:1,2
3.3
threshold cycle crossing point
point of the amplification curve at which the fluorescence signal rises above the baseline or crosses a
predefined threshold setting
[SOURCE: ISO 22119:2011, 3.17]
4 Principle
4.1 General
The identification of the monophasic variant of Salmonella Typhimurium comprises the three successive steps
descr
...

PROJET FINAL
Norme
internationale
ISO/FDIS 6579-4
ISO/TC 34/SC 9
Microbiologie de la chaîne
Secrétariat: AFNOR
alimentaire — Méthode
Début de vote:
horizontale pour la recherche, le
2024-10-31
dénombrement et le sérotypage des
Vote clos le:
Salmonella —
2024-12-26
Partie 4:
Identification du variant
monophasique de Salmonella
Typhimurium (1,4,[5],12:i:-) par
réaction de polymérisation en
chaîne (PCR)
Microbiology of the food chain — Horizontal method for the
detection, enumeration and serotyping of Salmonella —
Part 4: Identification of monophasic Salmonella Typhimurium
(1,4,[5],12:i:-) by polymerase chain reaction (PCR)
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
TRAITEMENT PARALLÈLE ISO/CEN
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
ISO/FDIS 6579-4:2024(fr) © ISO 2024

PROJET FINAL
ISO/FDIS 6579-4:2024(fr)
Norme
internationale
ISO/FDIS 6579-4
ISO/TC 34/SC 9
Microbiologie de la chaîne
Secrétariat: AFNOR
alimentaire — Méthode
Début de vote:
horizontale pour la recherche, le
2024-10-31
dénombrement et le sérotypage des
Vote clos le:
Salmonella —
2024-12-26
Partie 4:
Identification du variant
monophasique de Salmonella
Typhimurium (1,4,[5],12:i:-) par
réaction de polymérisation en
chaîne (PCR)
Microbiology of the food chain — Horizontal method for the
detection, enumeration and serotyping of Salmonella —
Part 4: Identification of monophasic Salmonella Typhimurium
(1,4,[5],12:i:-) by polymerase chain reaction (PCR)
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 2024 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
TRAITEMENT PARALLÈLE ISO/CEN
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
ISO/FDIS 6579-4:2024(fr) © ISO 2024

ii
ISO/FDIS 6579-4:2024(fr)
Sommaire Page
Avant-propos .iv
Introduction .v
1 Domaine d’application . 1
2 Références normatives . 2
3 Termes et définitions . 2
4 Principe. 3
4.1 Généralités .3
4.2 Préparation de colonies bien isolées .3
4.3 Mise en suspension d’une colonie .3
4.4 Amplification et détection . .3
5 Milieux de culture et réactifs . 3
6 Équipement et consommables . 3
7 Variant monophasique présumé de Salmonella Typhimurium . 4
8 Culture de l’isolat . 4
9 Mode opératoire . 4
9.1 Préparation de la suspension cellulaire ou de l’ADN .4
9.2 Amplification et détection par PCR .5
9.2.1 Généralités .5
9.2.2 Témoins de PCR .5
10 Expression des résultats . 5
11 Caractéristiques de performance des méthodes . 5
11.1 Validation conformément à l’ISO 17468 .5
11.2 Caractéristiques de performance .6
11.2.1 Étude d’évaluation de méthode(s) .6
11.2.2 Étude interlaboratoires .6
12 Rapport d’essai . 6
13 Assurance qualité . 7
Annexe A (normative) Milieux de culture et réactifs . 8
Annexe B (informative) Essai PCR multiplex en temps réel avec sonde pour l’identificationdu
variant monophasique de Salmonella Typhimurium (1,4,[5],12:i:-) .10
Annexe C (informative) Essai PCR multiplex avec électrophorèse sur gel d’agarose pour
l’identification du variant monophasique de Salmonella Typhimurium (1,4,[5],12:i:-) . 14
Annexe D (informative) Essai PCR simplex avec électrophorèse sur gel d’agarose pour
l’identification du variant monophasique de Salmonella Typhimurium (1,4,[5],12:i:-) .20
Annexe E (informative) Caractéristiques de performance .27
Bibliographie .34

iii
ISO/FDIS 6579-4:2024(fr)
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 propriété 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 brevet.
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 ISO/TC 34, Produits alimentaires, sous-comité SC 9,
Microbiologie., en collaboration avec le comité technique CEN/TC 463, Microbiologie de la chaîne alimentaire,
du Comité européen de normalisation (CEN), conformément à l’Accord de coopération technique entre l’ISO
et le CEN (Accord de Vienne).
Une liste de toutes les parties de la série ISO 6579 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.

iv
ISO/FDIS 6579-4:2024(fr)
Introduction
Plusieurs législations internationale, régionale et nationale fixent des limites réglementaires pour
garantir la soi-disant «absence» de Salmonella spp. dans des échantillons de la chaîne alimentaire. De plus,
plusieurs règlements de la Commission européenne (CE) prescrivent également l’absence de sérotypes
particuliers de Salmonella qui se sont révélés être à l’origine d’un pourcentage relativement important de
salmonelloses chez l’Homme. L’un de ces sérotypes de Salmonella pour lesquels des critères légaux ont
été fixés est Salmonella Typhimurium, y compris son variant monophasique 1,4,[5],12:i:- (par exemple,
[10]
Règlement (CE) n° 1086/2011 ). Il est donc important de savoir si un sérotype identifié ayant pour formule
antigénique 1,4,[5],12:i:- est bien le variant monophasique de Salmonella Typhimurium (1,4,[5],12:i:1,2) et
non le variant monophasique d’un autre sérotype de Salmonella (S.) pour lequel aucun critère n’a été fixé,
tel que S. Lagos (1,4,[5],12:i:1,5), S. Agama (4,12:i:1,6), S. Farsta (4,[5],12:i:e,n,x), S. Tsevie (1,4,12:i:e,n,z ),
S. Gloucester (1,4,12,27:i:l,w) ou S. Tumodi (1,4,12:i:z ). La confirmation de la distinction entre les sérotypes
de Salmonella Typhimurium et Salmonella non Typhimurium peut êtr
...