EN ISO 16654:2001/A1:2017

Microbiology of food and animal feeding stuffs - Horizontal method for the detection of Escherichia coli O157 - Amendment 1: Annex B: Result of interlaboratory studies (ISO 16654:2001/Amd 1:2017)

This standard describes the detection of E.coli O157 (Reference document EN ISO 16654)

Mikrobiologie von Lebensmitteln und Futtermitteln - Horizontales Verfahren für den Nachweis von Escherichia coli O157 - Änderung 1: Anhang B: Ergebnisse von Ringversuchen (ISO 16654:2001/Amd.1:2017)

Microbiologie des aliments - Méthode horizontale pour la recherche des Escherichia coli O157 - Amendement 1: Annexe B: Résultat des études interlaboratoires (ISO 16654:2001/Amd 1:2017)

Mikrobiologija živil in krme - Horizontalna metoda za ugotavljanje Escherichia coli O157 - Dopolnilo A1: Dodatek B: Rezultat medlaboratorijske študije (ISO 16654:2001/Amd 1:2017)

General Information

• Status: Published
• Publication Date: 25-Apr-2017
• Withdrawal Date: 30-Oct-2017
• ICS: 07.100.30 - Food microbiology
• Technical Committee: CEN/TC 463 - Microbiology of the food chain
• Drafting Committee: CEN/TC 463 - Microbiology of the food chain
• Current Stage: 6060 - Definitive text made available (DAV) - Publishing
• Start Date: 26-Apr-2017
• Due Date: 28-Feb-2014
• Completion Date: 26-Apr-2017
• Directive: 882/2004 - Regulation (EC) No 882/2004 of the European Parliament and of the Council of 29 April 2004 onofficial controls performed to ensure the verification of compliance with feed and food law, animal health and animal welfare rules
• Mandate: M/381 - Mandate for standardisation addressed to CEN in the field of Methods of analysis of foodstuffs concerning Food Hygiene
• Ref Project: SIST EN ISO 16654:2002/A1:2017 - Microbiology of food and animal feeding stuffs - Horizontal method for the detection of Escherichia coli O157 - Amendment 1: Annex B: Result of interlaboratory studies (ISO 16654:2001/Amd 1:2017)

Relations

• Amends: EN ISO 16654:2001 - Microbiology of food and animal feeding stuffs - Horizontal method for the detection of Escherichia coli O157 (ISO 16654:2001)
• Effective Date: 22-Dec-2008

Overview

EN ISO 16654:2001/A1:2017 is a key European standard focused on the microbiological analysis of food and animal feeding stuffs, specifically targeting the detection of Escherichia coli O157. This amendment updates the original EN ISO 16654:2001 by incorporating results from comprehensive interlaboratory studies to enhance the reliability and applicability of the horizontal detection method. Developed by CEN in collaboration with ISO technical committees, this standard addresses food safety concerns related to E. coli O157, a pathogenic strain posing serious health risks. The amendment also aligns with national regulations across multiple European countries and supports harmonized food testing practices.

Key Topics

• Horizontal detection method: Standardized protocol for identifying E. coli O157 in food and animal feed matrices, ensuring consistent and reproducible microbiological testing.
• Interlaboratory studies: Detailed results from international collaborative tests assessing the method’s performance characteristics - specificity, sensitivity, and limit of detection (LOD) - across various food types such as minced meat, raw milk, lettuce, and sprouts.
• Performance validation:
  • Sensitivity: The method’s ability to detect E. coli O157 in contaminated samples.
  • Specificity: Accuracy in confirming the absence of E. coli O157 in negative samples.
  • Limit of Detection (LOD): Defined as the concentration with a 50% probability of detection, with validated LOD for sprouts established at 8.4 cfu/g.
• Test portion flexibility: Adaptation allowances for test portion sizes while maintaining enrichment broth ratios, streamlining lab workflow without compromising test validity.
• Standardized reporting: Updated clauses for documenting test reports and method performance, ensuring clear communication of microbiological analysis outcomes.

Applications

• Food safety laboratories: Implements a validated and harmonized method for detecting E. coli O157 in diverse food and feed samples, critical for pathogen monitoring and compliance testing.
• Regulatory agencies: Provides a recognized testing framework utilized in official controls and risk assessment procedures to protect public health.
• Food industry quality assurance: Enables manufacturers to perform routine screening to prevent contamination and product recalls, particularly in sectors dealing with meat, dairy, fresh produce, and sprouted seeds.
• Research institutions: Supports method development and validation research for advancing microbiological detection techniques.
• International trade compliance: Facilitates mutual recognition of test results among European nations, reducing technical barriers and promoting food export safety standards.

Related Standards

• ISO 16140 (All parts): Provides guidelines for validation and verification of microbiological methods, referenced for validating test portions and pooling samples in EN ISO 16654.
• ISO 6887-1:2017: Specifies sample preparation methods for microbiological testing, used for verification protocols in pooling samples.
• NMKL N.164:1999: Another recognized method for E. coli O157 detection in food and feed, assessed as equivalent and integrated into the interlaboratory validation efforts.
• ISO 17025: Accreditation standard ensuring competence of laboratories performing this microbiological method.
• ISO/TS 19036: Technical specification used for computing measurement uncertainty in microbial detection tests.

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Keywords: EN ISO 16654:2001/A1:2017, Escherichia coli O157 detection, food microbiology standard, horizontal method, interlaboratory studies, microbiological analysis, sensitivity, specificity, limit of detection, food safety testing, pathogen detection in food, food and feed testing methods, CEN standard, microbiology of food, validation of microbiological methods.

Standards Content (Sample)

SLOVENSKI STANDARD
01-julij-2017
Mikrobiologija živil in krme - Horizontalna metoda za ugotavljanje Escherichia coli
O157 - Dopolnilo A1: Dodatek B: Rezultat medlaboratorijske študije (ISO
16654:2001/Amd 1:2017)
Microbiology of food and animal feeding stuffs - Horizontal method for the detection of
Escherichia coli O157 - Amendment 1: Annex B: Result of interlaboratory studies (ISO
16654:2001/Amd 1:2017)
Mikrobiologie von Lebensmitteln und Futtermitteln - Horizontales Verfahren für den
Nachweis von Escherichia coli O157 - Änderung 1 (ISO 16654:2001/Amd 1:2017)
Microbiologie des aliments - Méthode horizontale pour la recherche des Escherichia coli
O157 - Amendement 1: Annexe B: Résultat des études interlaboratoires (ISO
16654:2001/Amd 1:2017)
Ta slovenski standard je istoveten z: EN ISO 16654:2001/A1:2017
ICS:
07.100.30 Mikrobiologija živil Food microbiology
2003-01.Slovenski inštitut za standardizacijo. Razmnoževanje celote ali delov tega standarda ni dovoljeno.

EN ISO 16654:2001/A1
EUROPEAN STANDARD
NORME EUROPÉENNE
April 2017
EUROPÄISCHE NORM
ICS 07.100.30
English Version
Microbiology of food and animal feeding stuffs - Horizontal
method for the detection of Escherichia coli O157 -
Amendment 1: Annex B: Result of interlaboratory studies
(ISO 16654:2001/Amd 1:2017)
Microbiologie des aliments - Méthode horizontale pour Mikrobiologie von Lebensmitteln und Futtermitteln -
la recherche des Escherichia coli O157 - Amendement Horizontales Verfahren für den Nachweis von
1: Annexe B: Résultat des études interlaboratoires (ISO Escherichia coli O157 - Änderung 1 (ISO
16654:2001/Amd 1:2017) 16654:2001/Amd 1:2017)
This amendment A1 modifies the European Standard EN ISO 16654:2001; it was approved by CEN on 17 March 2017.

CEN members are bound to comply with the CEN/CENELEC Internal Regulations which stipulate the conditions for inclusion of
this amendment into the relevant 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 amendment 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, Former Yugoslav Republic of Macedonia, France, Germany, Greece, Hungary, Iceland, Ireland, Italy, Latvia, Lithuania,
Luxembourg, Malta, Netherlands, Norway, Poland, Portugal, Romania, Serbia, 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
© 2017 CEN All rights of exploitation in any form and by any means reserved Ref. No. EN ISO 16654:2001/A1:2017 E
worldwide for CEN national Members.

Contents Page
European foreword . 3

European foreword
This document (EN ISO 16654:2001/A1:2017) 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 Amendment to the European Standard EN ISO 16654:2001 shall be given the status of a national
standard, either by publication of an identical text or by endorsement, at the latest by October 2017,
and conflicting national standards shall be withdrawn at the latest by October 2017.
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.
This document has been prepared under a mandate given to CEN by the European Commission and the
European Free Trade Association.
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, Former Yugoslav Republic of Macedonia,
France, Germany, Greece, Hungary, Iceland, Ireland, Italy, Latvia, Lithuania, Luxembourg, Malta,
Netherlands, Norway, Poland, Portugal, Romania, Serbia, Slovakia, Slovenia, Spain, Sweden, Switzerland,
Turkey and the United Kingdom.
Endorsement notice
The text of ISO 16654:2001/Amd 1:2017 has been approved by CEN as EN ISO 16654:2001/A1:2017
without any modification.
INTERNATIONAL ISO
STANDARD 16654
First edition
2001-05-01
AMENDMENT 1
2017-03
Microbiology of food and animal
feeding stuffs — Horizontal method
for the detection of Escherichia coli
O157
AMENDMENT 1: Annex B: Result of
interlaboratory studies
Microbiologie des aliments — Méthode horizontale pour la recherche
des Escherichia coli O157
AMENDEMENT 1: Annexe B: Résultat des études interlaboratoires
Reference number
ISO 16654:2001/Amd.1:2017(E)
©
ISO 2017
ISO 16654:2001/Amd.1:2017(E)
© ISO 2017, Published in Switzerland
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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Tel. [phone]
Fax [phone]
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ii © ISO 2017 – All rights reserved

ISO 16654:2001/Amd.1:2017(E)
Foreword
ISO (the International Organization for Standardization) is a worldwide federation of national standards
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committee has been established has the right to be represented on that committee. International
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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 documents 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).
Attention is drawn to the possibility that some of the elements of this document may be the subject of
patent rights. ISO shall not be held responsible for identifying any or all such patent rights. Details of
any patent rights identified during the development of the document will be in the Introduction and/or
on the ISO list of patent declarations received (see www .iso .org/ patents).
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URL: w w w . i s o .org/ iso/ foreword .html.
This document was prepared by the European Committee for Standardization (CEN) Technical
Committee CEN/TC 275, Food analysis — Horizontal methods, in collaboration with ISO Technical
Committee TC 34, Food products, Subcommittee SC 9, Microbiology, in accordance with the agreement
on technical cooperation between ISO and CEN (Vienna Agreement).
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