1、BS EN ISO 6579-1:2017Microbiology of the food chain Horizontal method for thedetection, enumeration andserotyping of SalmonellaPart 1: Detection of Salmonella spp. (ISO6579-1:2017)BSI Standards PublicationWB11885_BSI_StandardCovs_2013_AW.indd 1 15/05/2013 15:06BS EN ISO 6579-1:2017 BRITISH STANDARDN
2、ational forewordThis British Standard is the UK implementation of EN ISO 6579-1:2017. It supersedes BS EN ISO 6785:2007 and BS EN ISO 6579:2002+A1:2007 which are withdrawn.The UK participation in its preparation was entrusted to Technical Committee AW/9, Microbiology.A list of organizations represen
3、ted on this committee can be obtained on request to its secretary.This publication does not purport to include all the necessary provisions of a contract. Users are responsible for its correct application. The British Standards Institution 2017.Published by BSI Standards Limited 2017ISBN 978 0 580 8
4、5340 1 ICS 07.100.30; 67.100.01 Compliance with a British Standard cannot confer immunity from legal obligations.This British Standard was published under the authority of the Standards Policy and Strategy Committee on 31 March 2017.Amendments/corrigenda issued since publicationDate Text affectedEUR
5、OPEAN STANDARD NORME EUROPENNE EUROPISCHE NORM EN ISO 6579-1 March 2017 ICS 07.100.30 Supersedes EN ISO 6579:2002, EN ISO 6785:2007English Version Microbiology of the food chain - Horizontal method for the detection, enumeration and serotyping of Salmonella - Part 1: Detection of Salmonella spp. (IS
6、O 6579-1:2017) Microbiologie de la chane alimentaire - Mthode horizontale pour la recherche, le dnombrement et le srotypage des Salmonella - Partie 1: Recherche des Salmonella spp. (ISO 6579-1:2017) Mikrobiologie der Lebensmittelkette - Horizontales Verfahren zum Nachweis, zur Zhlung und zur Serotyp
7、isierung von Salmonellen - Teil 1: Nachweis von Salmonella spp. (ISO 6579-1:2017) This European Standard was approved by CEN on 3 February 2017. 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
8、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
9、 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, Cypr
10、us, 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 Un
11、ited Kingdom. EUROPEAN COMMITTEE FOR STANDARDIZATION COMIT EUROPEN DE NORMALISATION EUROPISCHES KOMITEE FR 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 worldwide for CEN national Members. Ref. No. E
12、N ISO 6579-1:2017 EBS EN ISO 6579-1:2017EN ISO 6579-1:2017 (E) 3 European foreword This document (EN ISO 6579-1:2017) has been prepared by Technical Committee CEN/TC 275 “Food analysis - Horizontal methods”, the secretariat of which is held by DIN, in collaboration with Technical Committee ISO/TC 34
13、 “Food products“. 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 September 2017 and conflicting national standards shall be withdrawn at the latest by September 2017. Attention is drawn to the po
14、ssibility 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 supersedes EN ISO 6579:2002 and EN 6785:2007. This document has been prepared under a mandate given
15、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, De
16、nmark, 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. Endo
17、rsement notice The text of ISO 6579-1:2017 has been approved by CEN as EN ISO 6579-1:2017 without any modification. BS EN ISO 6579-1:2017ISO 6579-1:2017(E)Foreword vIntroduction vii1 Scope . 12 Normative references 13 Terms and definitions . 14 Principle 24.1 General . 24.2 Pre-enrichment in non-sel
18、ective liquid medium . 24.3 Enrichment in/on selective media . 24.4 Plating out on selective solid media . 24.5 Confirmation 35 Culture media, reagents, and antisera . 36 Equipment and consumables 37 Sampling 48 Preparation of test sample . 49 Procedure (see diagrams in Annex A) 49.1 Test portion an
19、d initial suspension . 49.2 Non-selective pre-enrichment . 49.3 Selective enrichment 59.3.1 General 59.3.2 Procedure for food, animal feed samples, and environmental samples from the food production area . 59.3.3 Procedure for samples from the primary production stage . 59.4 Plating out . 69.4.1 Gen
20、eral 69.4.2 Procedure for food, animal feed samples, and environmental samples from the food production area . 69.4.3 Procedure for samples from the primary production stage . 69.5 Confirmation . 79.5.1 General 79.5.2 Selection of colonies for confirmation 79.5.3 Biochemical testing . 89.5.4 Serolog
21、ical testing 119.5.5 Interpretation of biochemical and serological reactions .119.5.6 Serotyping. 1210 Expression of results .1211 Performance characteristics of the method 1211.1 Interlaboratory studies . 1211.2 Sensitivity . 1211.3 Specificity . 1211.4 LOD501212 Test report 13Annex A (normative) D
22、iagrams of the procedures .14Annex B (normative) Culture media and reagents .17Annex C (informative) Method validation studies and performance characteristics .32Annex D (normative) Detection of Salmonella enterica subspecies enterica serovars Typhi and Paratyphi 38 ISO 2017 All rights reserved iiiC
23、ontents PageBS EN ISO 6579-1:2017ISO 6579-1:2017(E)Annex E (informative) Examples of selective plating-out media 43Bibliography .48iv ISO 2017 All rights reservedBS EN ISO 6579-1:2017ISO 6579-1:2017(E)ForewordISO (the International Organization for Standardization) is a worldwide federation of natio
24、nal 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. Internati
25、onal 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
26、 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 .is
27、o .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 wi
28、ll be in the Introduction and/or on the ISO list of patent declarations received (see www .iso .org/ patents).Any trade name used in this document is information given for the convenience of users and does not constitute an endorsement.For an explanation on the meaning of ISO specific terms and expr
29、essions related to conformity assessment, as well as information about ISOs adherence to the World Trade Organization (WTO) principles in the Technical Barriers to Trade (TBT) see the following URL: www .iso .org/ iso/ foreword .htmlThis document was prepared by the European Committee for Standardiz
30、ation (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).This first edition of I
31、SO 6579-1 cancels and replaces ISO 6579:2002 and ISO 6785:2001, which have been technically revised. It also incorporates ISO 6579:2002/Amd 1:2007 and ISO 6579:2002/Cor 1:2004. The main changes, compared to ISO 6579:2002, are the following. ISO 6785 has been incorporated in this document. Samples fr
32、om the primary production stage have been added to the scope. Detection of Salmonella Typhi and Salmonella Paratyphi is described in Annex D. Descriptions of preparations of initial suspensions have been removed and references made to relevant parts of ISO 6887, whenever possible. The temperature ra
33、nge for incubation of non-selective media has been extended from 37 C 1 C to 34 C to 38 C without further tolerance. For selective enrichment, there is a choice between using the broth or the semi-solid agar of Rappaport Vassiliadis medium (RVS or MSRV) for food, animal feed samples, and for environ
34、mental samples from the food production area. The inoculation of the isolation medium has become less prescriptive; the objective is to obtain well-isolated colonies after incubation. For confirmation, it is acceptable to perform the tests on only one suspect colony (instead of one suspect colony of
35、 each medium combination). If this isolate tests negative for Salmonella, four more suspect isolates from different media combinations shall be tested. ISO 2017 All rights reserved vBS EN ISO 6579-1:2017ISO 6579-1:2017(E) It is permitted to perform the biochemical confirmation directly on a suspect,
36、 well-isolated colony from the selective plating medium. The purity check on the non-selective agar medium can then be performed in parallel. Two confirmation tests have become optional (-galactosidase test and indole reaction) and one confirmation test has been deleted (Voges-Proskauer reaction). I
37、n this document, serological confirmation (to serogroup level) is described. For guidance on serotyping (to serovar level), reference is made to ISO/TR 6579-3. Table 1 has been improved. Performance testing for the quality assurance of the culture media has been added to Annex B. Performance charact
38、eristics of MSRV have been added to Annex C.A list of all parts in the ISO 6579 series can be found on the ISO website.vi ISO 2017 All rights reservedBS EN ISO 6579-1:2017ISO 6579-1:2017(E)IntroductionThis document describes a horizontal method for the detection of Salmonella spp. in food (including
39、 milk and milk products, originally described in ISO 6785), in animal feed, in animal faeces, and in environmental samples from the primary production stage (the latter two were originally described in ISO 6579:2002/Amd 1:2007).The main changes, listed in the foreword, introduced in this document co
40、mpared to ISO 6579:2002, are considered as minor (see ISO 1746837).A procedure for the enumeration of Salmonella spp. is described in ISO/TS 6579-2.3Guidance for serotyping of Salmonella spp. is described in ISO/TR 6579-3.24 ISO 2017 All rights reserved viiBS EN ISO 6579-1:2017BS EN ISO 6579-1:2017M
41、icrobiology of the food chain Horizontal method for the detection, enumeration and serotyping of Salmonella Part 1: Detection of Salmonella spp.WARNING In order to safeguard the health of laboratory personnel, it is essential that tests for detecting Salmonella are only undertaken in properly equipp
42、ed 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, associat
43、ed with its use. It is the responsibility of the user to establish appropriate safety and health practices and to ensure compliance with any national regulatory conditions.1 ScopeThis document specifies a horizontal method for the detection of Salmonella. It is applicable to the following: products
44、intended for human consumption and the feeding of animals; environmental samples in the area of food production and food handling; samples from the primary production stage such as animal faeces, dust, and swabs.With this horizontal method, most of the Salmonella serovars are intended to be detected
45、. For the detection of some specific serovars, additional culture steps may be needed. For Salmonella Typhi and Salmonella Paratyphi, the procedure is described in Annex D.The selective enrichment medium modified semi-solid Rappaport-Vassiliadis (MSRV) agar is intended for the detection of motile Sa
46、lmonella and is not appropriate for the detection of non-motile Salmonella strains.2 Normative referencesThe 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
47、. For undated references, the latest edition of the referenced document (including any amendments) applies.ISO 6887 (all parts), Microbiology of food and animal feed Preparation of test samples, initial suspension and decimal dilutions for microbiological examinationISO 7218, Microbiology of food an
48、d animal feeding stuffs General requirements and guidance for microbiological examinationsISO 11133:2014, Microbiology of food, animal feed and water Preparation, production, storage and performance testing of culture media3 Terms and definitionsFor the purposes of this document, the following terms
49、 and definitions apply.INTERNATIONAL STANDARD ISO 6579-1:2017(E) ISO 2017 All rights reserved 1BS EN ISO 6579-1:2017ISO 6579-1:2017(E)ISO and IEC maintain terminological databases for use in standardization at the following addresses: IEC Electropedia: available at h t t p :/ www .electropedia .org/ ISO Online browsing platform: available at h t t p :/ www .iso .org/ obp3.1Salmonellamicroorganism which forms typical or less typical colonies on solid selective media and which displays the characteristics de
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