SIST EN ISO 6579-4:2025
(Main)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) (ISO 6579-4:2025)
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) (ISO 6579-4:2025)
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.
Mikrobiologie der Lebensmittelkette - Horizontales Verfahren zum Nachweis von Salmonella spp. - Teil 4: Identifizierung von monophasischen Salmonella Typhimurium (1,4,[5],12:i:-) durch Polymerase‑Kettenreaktion (PCR) (ISO 6579-4:2025)
Dieses Dokument legt ein horizontales In-vitro-Verfahren für die molekulare Identifizierung und Differenzierung der monophasischen Variante von Salmonella enterica subsp. enterica serovar Typhimurium (1,4[5],12:i: ), bei der die zweite H-Phase H:1,2 fehlt, und mit den Isolaten beginnt fest. Dieses Verfahren weist spezifische DNA-Sequenzen einer intergenischen Region der ersten H-Phase des Flagellen-Clusters nach, um Salmonella enterica subsp. enterica serovar Typhimurium (im folgenden Salmonella Typhimurium) sowie spezifische DNA-Sequenzen von Genen, die mit der zweiten H-Phase der Flagellen-Antigen-Expression verbunden sind, zu identifizieren.
Das Verfahren ist anwendbar zur:
Differenzierung des untersuchten Isolats zwischen monophasischem Salmonella Typhimurium und der monophasischen Variante eines anderen Salmonella non Typhimurium-Serovars, das die gleiche Antigenformel hat;
Identifizierung des untersuchten Isolats, das entweder ein monophasisches Salmonella Typhimurium oder ein (biphasisches) Salmonella Typhimurium ist.
Dieses Dokument ist anwendbar auf die Untersuchung einer zur Gattung Salmonellen gehörenden Reinkultur, welche isoliert wurde von:
Produkten, die für den menschlichen Verzehr vorgesehen sind;
Produkten, die als Futtermittel vorgesehen sind;
Umgebungsproben im Bereich der Herstellung und Handhabung von Lebensmitteln und Futtermitteln;
Proben aus dem Bereich der Primärproduktion.
Dieses Dokument kann auch in anderen Bereichen (z. B. Umwelt, Gesundheit von Menschen, Gesundheit von Tieren) zur Identifizierung von monophasischem Salmonella Typhimurium angewandt werden.
ANMERKUNG Dieses Verfahren wurde in einer Studie zur Verfahrensbewertung sowie in einem Ringversuch mit einem umfangreichen Satz unterschiedlicher Stämme (Ziel- und Nicht-Ziel-Stämme) validiert, die aus verschiedenen Quellen (Lebensmittelerzeugnisse, Tiere, Futtermittel, Primärproduktionsproben und Menschen) isoliert wurden. Detaillierte Informationen zur Validierung siehe Anhang E.
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) (ISO 6579-4:2025)
Le présent document spécifie une méthode horizontale in vitro d’identification moléculaire et de différenciation du variant monophasique de Salmonella enterica sous-espèce enterica sérotype Typhimurium (1,4,[5],12:i:-) ne présentant pas l’antigène H de seconde phase H:1,2, à partir d’isolats. La méthode détecte des séquences d’ADN spécifiques d’une région intergénique du groupe des flagellines de première phase H pour l’identification de Salmonella enterica sous-espèce enterica sérotype Typhimurium (encore appelé Salmonella Typhimurium) et des séquences d’ADN spécifiques de gènes associés à l’expression d’antigène flagellaire H de seconde phase.
La méthode est applicable pour:
— la différenciation de l’isolat analysé entre le variant monophasique de Salmonella Typhimurium et le variant monophasique d’un autre sérotype de Salmonella non Typhimurium ayant la même formule antigénique;
— l’identification de l’isolat analysé comme étant soit le variant monophasique de Salmonella Typhimurium, soit Salmonella Typhimurium (biphasique).
Le présent document est applicable à l’analyse d’une culture pure appartenant au genre Salmonella, isolée à partir:
— de produits destinés à la consommation humaine;
— de produits destinés à l’alimentation animale;
— d’échantillons environnementaux dans le domaine de la production et de la manutention de denrées alimentaires;
— d’échantillons au stade de la production primaire.
Le présent document peut aussi être appliqué dans d’autres domaines pour l’identification du variant monophasique de Salmonella Typhimurium (par exemple, domaine environnemental, santé humaine, santé animale).
NOTE La présente méthode a été validée dans la cadre d’une étude d’évaluation de méthode et d’une étude interlaboratoires à l’aide d’un large panel de souches différentes (souches cibles et non cibles), isolées à partir de différentes sources (produits alimentaires, animaux, aliments pour animaux, échantillons de production primaire et échantillons humains). Pour de plus amples informations sur la validation, voir Annexe E.
Mikrobiologija v prehranski verigi - Horizontalna metoda za ugotavljanje prisotnosti, štetja in serotipizacije Salmonella - 4. del: Identifikacija monofazne Salmonella Typhimurium (1,4,[5],12:i:-) s polimerazno verižno reakcijo (PCR) (ISO 6579-4:2025)
General Information
Standards Content (Sample)
SLOVENSKI STANDARD
01-april-2025
Mikrobiologija v prehranski verigi - Horizontalna metoda za ugotavljanje
prisotnosti, štetja in serotipizacije Salmonella - 4. del: Identifikacija monofazne
Salmonella Typhimurium (1,4,[5],12:i:-) s polimerazno verižno reakcijo (PCR) (ISO
6579-4:2025)
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) (ISO 6579-4:2025)
Mikrobiologie der Lebensmittelkette - Horizontales Verfahren zum Nachweis von
Salmonella spp. - Teil 4: Identifizierung von monophasischen Salmonella Typhimurium
(1,4,[5],12:i:-) durch Polymerase‑Kettenreaktion (PCR) (ISO 6579-4:2025)
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) (ISO 6579-4:2025)
Ta slovenski standard je istoveten z: EN ISO 6579-4:2025
ICS:
07.100.30 Mikrobiologija živil Food microbiology
2003-01.Slovenski inštitut za standardizacijo. Razmnoževanje celote ali delov tega standarda ni dovoljeno.
EN ISO 6579-4
EUROPEAN STANDARD
NORME EUROPÉENNE
February 2025
EUROPÄISCHE NORM
ICS 07.100.30
English Version
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) (ISO
6579-4:2025)
Microbiologie de la chaîne alimentaire - Méthode Mikrobiologie der Lebensmittelkette - Horizontales
horizontale pour la recherche, le dénombrement et le Verfahren zum Nachweis, zur Zählung und zur
sérotypage des Salmonella - Partie 4: Identification du Serotypisierung von Salmonellen - Teil 4:
variant monophasique de Salmonella Typhimurium Identifizierung von monophasischen Salmonella
(1,4,[5],12:i:-) par réaction de polymérisation en Typhimurium (1,4,[5],12:i:-) durch Polymerase-
chaîne (PCR) (ISO 6579-4:2025) Kettenreaktion (PCR) (ISO 6579-4:2025)
This European Standard was approved by CEN on 27 January 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 6579-4:2025 E
worldwide for CEN national Members.
Contents Page
European foreword . 3
European foreword
This document (EN ISO 6579-4:2025) 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 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 August 2025, and conflicting national standards shall
be withdrawn at the latest by August 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.
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 6579-4:2025 has been approved by CEN as EN ISO 6579-4:2025 without any
modification.
International
Standard
ISO 6579-4
First edition
Microbiology of the food chain —
2025-01
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)
Reference number
ISO 6579-4:2025(en) © ISO 2025
ISO 6579-4:2025(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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Published in Switzerland
ii
ISO 6579-4:2025(en)
Contents Page
Foreword .iv
Introduction .v
1 Scope . 1
2 Normative references . 2
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
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 6579-4:2025(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 ca
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