CEN ISO/TS 17919:2013

Microbiology of the food chain - Polymerase chain reaction (PCR) for the detection of food-borne pathogens - Detection of botulinum type A, B, E and F neurotoxin-producing clostridia (ISO/TS 17919:2013)

ISO/TS 17919:2013 specifies a horizontal method for the molecular detection of clostridia carrying botulinum neurotoxin A, B, E, and F genes by a PCR method. This method detects the genes and not the toxins, therefore a positive result does not necessarily mean the presence of these toxins in the sample investigated. ISO/TS 17919:2013 is applicable to products for human consumption, animal feed, and environmental samples.
The PCR assays for detection of genetic sequences encoding specific toxin types are described in annexes.

Mikrobiologie der Lebensmittelkette - Polymerase-Kettenreaktion (PCR) zum Nachweis von pathogenen Mikroorganismen in Lebensmitteln - Nachweis von Botulinum-Neurotoxin Typ A, B, E und F produzierenden Clostridien (ISO/TS 17919:2013)

Microbiologie de la chaîne alimentaire - Réaction de polymérisation en chaîne (PCR) pour la détection de micro-organismes pathogènes dans les aliments - Détection des clostridies productrices de neurotoxine botulique de type A, B, E et F (ISO/TS 17919:2013)

L'ISO/TS 17919:2013 spécifie une méthode horizontale de détection moléculaire des clostridies portant les gènes A, B, E et F de neurotoxine botulique par réaction de polymérisation en chaîne (PCR). Cette méthode détecte les gènes, et non les toxines. Par conséquent, un résultat positif n'est pas nécessairement synonyme de présence de ces toxines dans l'échantillon examiné. L'ISO/TS 17919:2013 s'applique aux produits destinés à être consommés par l'Homme, aux aliments pour animaux et aux échantillons environnementaux.
Les réactions de PCR de détection des séquences génétiques codant ces types particuliers de toxine sont décrites.

Mikrobiologija v prehranski verigi - Polimerazna verižna reakcija (PCR) za ugotavljanje prisotnosti patogenih mikroorganizmov v živilih - Ugotavljanje prisotnosti klostridijev, ki tvorijo botulinusne nevrotoksine tipov A, B, E in F (ISO/TS 17919:2013)

Ta tehnična specifikacija določa horizontalno metodo za molekularno ugotavljanje prisotnosti klostridijev, ki prenašajo botulinusne nevrotoksine genov A, B, E in F, z metodo polimerazne verižne reakcije (PCR). S to metodo se ugotavlja prisotnost genov in ne toksinov, zato pozitiven rezultat ne pomeni nujno prisotnosti teh toksinov v preiskovanem vzorcu. Ta tehnična specifikacija se uporablja za proizvode za prehrano ljudi, živalsko krmo in okoljske vzorce. Preskusi PCR za ugotavljanje prisotnosti genskih zaporedij specifičnih tipov toksinov so opisani v dodatkih B in C.

General Information

• Status: Published
• Publication Date: 05-Nov-2013
• ICS: 07.100.30 - Food microbiology
• Technical Committee: CEN/TC 463 - Microbiology of the food chain
• Drafting Committee: CEN/TC 463/WG 1 - General requirements relating to PCR methods
• Current Stage: 6060 - Definitive text made available (DAV) - Publishing
• Start Date: 06-Nov-2013
• Due Date: 24-Apr-2013
• Completion Date: 06-Nov-2013
• Ref Project: SIST-TS CEN ISO/TS 17919:2014 - Microbiology of the food chain - Polymerase chain reaction (PCR) for the detection of food-borne pathogens - Detection of botulinum type A, B, E and F neurotoxin-producing clostridia (ISO/TS 17919:2013)

Standards Content (Sample)

SLOVENSKI STANDARD
01-januar-2014
Mikrobiologija v prehranski verigi - Polimerazna verižna reakcija (PCR) za
ugotavljanje prisotnosti patogenih mikroorganizmov v živilih - Ugotavljanje
prisotnosti klostridijev, ki tvorijo botulinusne nevrotoksine tipov A, B, E in F
(ISO/TS 17919:2013)
Microbiology of the food chain - Polymerase chain reaction (PCR) for the detection of
food-borne pathogens - Detection of botulinum type A, B, E and F neurotoxin-producing
clostridia (ISO/TS 17919:2013)
Mikrobiologie von Lebensmitteln, Futtermitteln und Umgebungsproben - Polymerase-
Kettenreaktion (PCR) zum Nachweis von pathogenen Mikroorganismen in Lebensmitteln
- Nachweis von Botulinum-Neurotoxin-Typ A, B, E und F produzierenden Clostridien
(ISO/TS 17919:2013)
Microbiologie de la chaîne alimentaire - Réaction de polymérisation en chaîne (PCR)
pour la détection de micro-organismes pathogènes dans les aliments - Détection des
clostridies productrices de neurotoxine botulique de type A, B, E et F (ISO/TS
17919:2013)
Ta slovenski standard je istoveten z: CEN ISO/TS 17919:2013
ICS:
07.100.30 Mikrobiologija živil Food microbiology
67.020 Procesi v živilski industriji Processes in the food
industry
2003-01.Slovenski inštitut za standardizacijo. Razmnoževanje celote ali delov tega standarda ni dovoljeno.

TECHNICAL SPECIFICATION
CEN ISO/TS 17919
SPÉCIFICATION TECHNIQUE
TECHNISCHE SPEZIFIKATION
November 2013
ICS 07.100.30
English Version
Microbiology of the food chain - Polymerase chain reaction
(PCR) for the detection of food-borne pathogens - Detection of
botulinum type A, B, E and F neurotoxin-producing clostridia
(ISO/TS 17919:2013)
Microbiologie de la chaîne alimentaire - Réaction de Mikrobiologie von Lebensmitteln, Futtermitteln und
polymérisation en chaîne (PCR) pour la détection de micro- Umgebungsproben - Polymerase-Kettenreaktion (PCR)
organismes pathogènes dans les aliments - Détection des zum Nachweis von pathogenen Mikroorganismen in
clostridies productrices de neurotoxine botulique de type A, Lebensmitteln - Nachweis von Botulinum-Neurotoxin-Typ A,
B, E et F (ISO/TS 17919:2013) B, E und F produzierenden Clostridien (ISO/TS
17919:2013)
This Technical Specification (CEN/TS) was approved by CEN on 16 April 2013 for provisional application.

The period of validity of this CEN/TS is limited initially to three years. After two years the members of CEN will be requested to submit their
comments, particularly on the question whether the CEN/TS can be converted into a European Standard.

CEN members are required to announce the existence of this CEN/TS in the same way as for an EN and to make the CEN/TS available
promptly at national level in an appropriate form. It is permissible to keep conflicting national standards in force (in parallel to the CEN/TS)
until the final decision about the possible conversion of the CEN/TS into an EN is reached.

CEN members are the national standards bodies of Austria, Belgium, Bulgaria, Croatia, Cyprus, Czech Republic, Denmark, Estonia,
Finland, Former Yugoslav Republic of Macedonia, France, Germany, Greece, Hungary, Iceland, Ireland, Italy, Latvia, Lithuania,
Luxembourg, Malta, Netherlands, Norway, Poland, Portugal, Romania, Slovakia, Slovenia, Spain, Sweden, Switzerland, Turkey and United
Kingdom.
EUROPEAN COMMITTEE FOR STANDARDIZATION
COMITÉ EUROPÉEN DE NORMALISATION

EUROPÄISCHES KOMITEE FÜR NORMUNG

CEN-CENELEC Management Centre: Avenue Marnix 17, B-1000 Brussels
© 2013 CEN All rights of exploitation in any form and by any means reserved Ref. No. CEN ISO/TS 17919:2013 E
worldwide for CEN national Members.

Contents Page
Foreword .3

Foreword
This document (CEN ISO/TS 17919:2013) 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.
Attention is drawn to the possibility that some of the elements of this document may be the subject of patent
rights. CEN [and/or CENELEC] shall not be held responsible for identifying any or all such patent rights.
According to the CEN-CENELEC Internal Regulations, the national standards organizations of the following
countries are bound to announce this Technical Specification: Austria, Belgium, Bulgaria, Croatia, Cyprus,
Czech Republic, Denmark, Estonia, Finland, Former Yugoslav Republic of Macedonia, France, Germany,
Greece, Hungary, Iceland, Ireland, Italy, Latvia, Lithuania, Luxembourg, Malta, Netherlands, Norway, Poland,
Portugal, Romania, Slovakia, Slovenia, Spain, Sweden, Switzerland, Turkey and the United Kingdom.
Endorsement notice
The text of ISO/TS 17919:2013 has been approved by CEN as CEN ISO/TS 17919:2013 without any
modification.
TECHNICAL ISO/TS
SPECIFICATION 17919
First edition
2013-11-01
Microbiology of the food chain —
Polymerase chain reaction (PCR) for
the detection of food-borne pathogens
— Detection of botulinum type A, B, E
and F neurotoxin-producing clostridia
Microbiologie de la chaîne alimentaire — Réaction de polymérisation
en chaîne (PCR) pour la détection de micro-organismes pathogènes
dans les aliments — Détection des clostridies productrices de
neurotoxine botulique de type A, B, E et F
Reference number
ISO/TS 17919:2013(E)
©
ISO 2013
ISO/TS 17919:2013(E)
© ISO 2013
All rights reserved. Unless otherwise specified, 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.
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ii © ISO 2013 – All rights reserved

ISO/TS 17919:2013(E)
Contents Page
Foreword .iv
Introduction .v
1 Scope . 1
2 Normative references . 1
3 Terms and definitions . 1
4 Symbols and abbreviated terms . 1
4.1 Symbols . 1
4.2 Abbreviated terms . 2
5 Principle . 2
5.1 General . 2
5.2 Microbial enrichment . 2
5.3 Nucleic acid extraction . 2
5.4 Amplification by PCR . 2
5.5 Detection of PCR products . 2
5.6 Confirmation . 2
6 Reagents . 3
6.1 General . 3
6.2 Culture media . 3
6.3 Nucleic acid extraction . 4
6.4 Reagents for PCR . 5
7 Apparatus and equipment . 5
7.1 General . 5
7.2 Equipment for sample preparation prior to enrichment . 5
7.3 Equipment for microbial enrichment . . 5
7.4 Equipment used for nucleic acid extraction . 6
7.5 Equipment used for PCR . 6
7.6 Equipment used for the detection of the PCR product . 6
8 Sampling . 7
9 Procedure. 7
9.1 Sample preparation prior to enrichment . 7
9.2 Microbial enrichment . 7
9.3 Nucleic acid preparation. 8
9.4 PCR amplification . 9
9.5 Confirmation of a positive PCR result . 9
Annex A (normative) Flow-diagram of the procedure .10
Annex B (informative) Multiplex PCR assays for detection of genes encoding botulinum
neurotoxin types A, B, E, and F using agarose gel electrophoresis .11
Annex C (informative) Assays for detection of genes encoding botulinum neurotoxin types A, B, E,
and F using real-time PCR .29
Annex D (informative) Preparation of C. botulinum spores.40
Bibliography .46
© ISO
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