1、BSI Standards Publication PD CEN ISO/TR 6579-3:2014 Microbiology of the food chain Horizontal method for the detection, enumeration and serotyping of Salmonella Part 3: Guidelines for serotyping of Salmonella spp. (ISO/TR 6579-3:2014) PD CEN ISO/TR 6579-3:2014 Incorporating corrigendum August 2014PD
2、 CEN ISO/TR 6579-3:2014 PUBLISHED DOCUMENT National foreword This Published Document is the UK implementation of CEN ISO/TR 6579-3:2014. The UK participation in its preparation was entrusted to Technical Committee AW/9, Microbiology. A list of organizations represented on this committee can be obtai
3、ned 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 2014. Published by BSI Standards Limited 2014 ISBN 978 0 580 87762 9 ICS 07.100.30 Compliance
4、 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 July 2014. Amendments/corrigenda issued since publication Date Text affected 31 August 2014 Implementation of CEN Corre
5、ction Notice 06 August 2014: supersession information removed from CEN title page and foreword. Supersession information removed from national foreword TECHNICAL REPORT RAPPORT TECHNIQUE TECHNISCHER BERICHT CEN ISO/TR 6579-3 July 2014 ICS 07.100.30 English Version Microbiology of the food chain - Ho
6、rizontal method for the detection, enumeration and serotyping of Salmonella - Part 3: Guidelines for serotyping of Salmonella spp. (ISO/TR 6579- 3:2014) Microbiologie de la chane alimentaire - Mthode horizontale pour la recherche, le dnombrement et la srotypie des Salmonella - Partie 3: Lignes direc
7、trices pour la srotypie des Salmonella spp. (ISO/TR 6579-3:2014) Mikrobiologie von Lebensmitteln und Futtermitteln - Horizontales Verfahren zum Nachweis, zur Zhlung und zur Serotypisierung von Salmonellen - Teil 3: Leitfaden fr die Serotypisierung von Salmonella spp. (ISO/TR 6579-3:2014)This Technic
8、al Report was approved by CEN on 28 October 2013. It has been drawn up by the Technical Committee CEN/TC 275. CEN members are the national standards bodies of Austria, Belgium, Bulgaria, Croatia, Cyprus, Czech Republic, Denmark, Estonia, Finland, Former Yugoslav Republic of Macedonia, France, German
9、y, 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 COMIT EUROPEN DE NORMALISATION EUROPISCHES KOMITEE FR N
10、ORMUNG CEN-CENELEC Management Centre: Avenue Marnix 17, B-1000 Brussels 2014 CEN All rights of exploitation in any form and by any means reserved worldwide for CEN national Members. Ref. No. CEN ISO/TR 6579-3:2014 EPD CEN ISO/TR 6579-3:2014 CEN ISO/TR 6579-3:2014 (E) 3 Foreword This document (CEN IS
11、O/TR 6579-3:2014) 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
12、 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. Endorsement notice The text of ISO/TR 6579-3:2014 has been approved by CEN as CEN ISO/TR 6579-3:2014 without any modification.
13、CEN ISO/TR 6579-3:2014 (E) 3 Foreword This document (CEN ISO/TR 6579-3:2014) 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
14、 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. Endorsement notice The text of ISO/TR 6579-3:2014 has been approved by CEN as CEN ISO/TR 6579-3:2014 without an
15、y modification. PD CEN ISO/TR 6579-3:2014 CEN ISO/TR 6579-3:2014 (E) 3 Foreword This document (CEN ISO/TR 6579-3:2014) 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
16、 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. This document supersedes EN ISO 6579:2002. Endorsement notice The tex
17、t of ISO/TR 6579-3:2014 has been approved by CEN as CEN ISO/TR 6579-3:2014 without any modification. PD CEN ISO/TR 6579-3:2014ISO/TR 6579-3:2014(E) ISO 2014 All rights reserved iii Contents Page Foreword iv Introduction v 1 Scope . 1 2 Normative references 1 3 T erms and definitions . 1 4 Principle
18、2 5 Culture media and sera 2 5.1 General . 2 5.2 Culture media and reagents . 2 5.3 Antisera . 2 6 Apparatus . 2 7 Sample . 3 8 Taxonomy of Salmonella . 3 8.1 General . 3 8.2 Nomenclature. 3 8.3 Biochemical characteristics . 4 8.4 Antigenic characteristics 5 9 Procedure for Salmonella serotyping . 7
19、 9.1 General . 7 9.2 Example procedure for serotyping five public health-related Salmonella serovars . 7 10 Quality control .11 11 Reporting .11 Annex A (informative) Composition and preparation of culture media and reagents 13 Annex B (informative) Examples of procedures for serotyping an unknown S
20、almonella isolate 20 Annex C (informative) Biochemical tests 24 Annex D (informative) S chematic o v ervie w for ser otyping fi v e important public-health r elat ed Salmonella serovars .26 Annex E (informative) Microtitre plate method for serotyping Salmonella spp27 Annex F (informative) Examples o
21、f procedures for phase inversion 29 Bibliography .32PD CEN ISO/TR 6579-3:2014ISO/TR 6579-3:2014(E) 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 c
22、arried 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 wor
23、k. 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 diff
24、erent 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 m
25、ay 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/pate
26、nts). 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 expressions related to conformity assessment, as well as information about ISOs adherence to the WTO princi
27、ples in the Technical Barriers to Trade (TBT) see the following URL: Foreword - Supplementary information The committee responsible for this document is ISO/TC 34, Food products, Subcommittee SC 9, Microbiology. ISO 6579 consists of the following parts, under the general title Microbiology of the fo
28、od chain Horizontal method for the detection, enumeration and serotyping of Salmonella: Part 1: Horizontal method for the detection of Salmonella spp. 1) Part 2: Enumeration by a miniaturized most probable number technique Technical Specification 2) Part 3: Guidelines for serotyping of Salmonella sp
29、p. Technical Report 1) Under preparation. (Revision of ISO 6579:2002) 2) The main element of the series title has been changed since Part 2 was published. It is intended that upon revision, the main element of the title will be aligned with Part 3.iv ISO 2014 All rights reservedPD CEN ISO/TR 6579-3:
30、2014ISO/TR 6579-3:2014(E) Introduction This part of ISO 6579 gives information on the taxonomy of Salmonella spp. and it gives guidance on serotyping of Salmonella serovars, based on the WhiteKauffmannLe Minor scheme (see Reference 9). ISO 2014 All rights reserved vPD CEN ISO/TR 6579-3:2014PD CEN IS
31、O/TR 6579-3:2014Microbiology of the food chain Horizontal method for the detection, enumeration and serotyping of Salmonella Part 3: Guidelines for serotyping of Salmonella spp. WARNING In order to safeguard the health of laboratory personnel, it is essential that tests for detecting and typing Salm
32、onella, be undertaken only in properly equipped laboratories, under the control of a skilled microbiologist, and that great care is taken in the disposal of all incubated materials. IMPORTANT Persons using this document should be familiar with normal laboratory practice. This document does not purpo
33、rt 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 and to ensure compliance with any national regulatory conditions. 1 Scope This part of ISO 6579 gives guidance on the procedure for sero
34、typing Salmonella serovars and is applicable to the serotyping of pure cultures of Salmonella spp., independent of the source from which they are isolated. 2 Normative references The following documents, in whole or in part, are normatively referenced in this document and are indispensable for its a
35、pplication. For dated references, only the edition cited applies. For undated references, the latest edition of the referenced document (including any amendments) applies. ISO 6579-1, Microbiology of the food chain Horizontal method for the detection, enumeration and serotyping of Salmonella Part 1:
36、 Horizontal method for the detection of Salmonella spp. ISO 7218, Microbiology of food and animal feeding stuffs General requirements and guidance for microbiological examinations ISO 11133, Microbiology of food, animal feed and water Preparation, production, storage and performance testing of cultu
37、re media 3 T erms a nd definiti ons For the purposes of this document, the following terms and definitions apply. 3.1 Salmonella gram-negative, oxidase-negative, facultatively anaerobic, non-spore-forming, rod-shaped bacteria which generally form colonies of 2 mm to 4 mm in diameter on solid selecti
38、ve media and display biochemical and serological characteristics described when tests are carried out in accordance with this part of ISO 6579 TECHNICAL REPORT ISO/TR 6579-3:2014(E) ISO 2014 All rights reserved 1PD CEN ISO/TR 6579-3:2014ISO/TR 6579-3:2014(E) 3.2 serotyping of Salmonella determinatio
39、n of the presence or absence of specific O-antigens, H-antigens and Vi-antigens in an isolate confirmed as Salmonella (3.1) 3.3 antigenic formula combination of numbers and letters representing the O-, H-, and Vi-antigens of an isolate confirmed as Salmonella (3.1) 4 Principle For the serotyping of
40、Salmonella spp. the following antigens are determined for isolates biochemically confirmed as Salmonella spp.: O-antigens, H-antigens and Vi-antigens. NOTE Alternative procedures can be used to confirm the isolate being Salmonella spp. provided the suitabilit y of the alternative procedure is verifi
41、ed (see ISO 7218). 5 Culture media and sera 5.1 General For current laboratory practice, apply ISO 7218. For the performance testing of media, follow the recommendations of ISO 11133. 5.2 Culture media and reagents See Annex A. 5.3 Antisera O-antisera, H-antisera and Vi-antisera are available from v
42、arious commercial suppliers. Information on relevant polyvalent antisera and monovalent antisera can be found in Annex B. 6 Apparatus Disposable supplies are an acceptable alternative to reusable glassware if they have similar specifications. Usual microbiological laboratory equipment and, in partic
43、ular, the following. 6.1 Incubator, to grow Salmonella isolates, capable of operating in the range 34 C to 38 C. NOTE In this part of ISO 6579, the incubation temperature is not a differential parameter. Isolates are cultured to obtain sufficient material to perform the tests on a pure culture. Ther
44、efore, culture step is performed at an optimal growth temperature. For Salmonella this is generally a temperature between 34 C and 38 C. 6.2 Oven (for dry sterilization) or autoclave (for wet sterilization). See ISO 7218. 6.3 Refrigerator (for storage of prepared media), capable of operating at 5 C
45、3 C. 6.4 Glass slides. 6.5 Sterile inoculation instrument, e.g. needles, wires, wooden sticks, loops (e.g. of 1 l).2 ISO 2014 All rights reservedPD CEN ISO/TR 6579-3:2014ISO/TR 6579-3:2014(E) 6.6 Sterile test tubes and flasks, of appropriate capacity. Flasks or bottles and test tubes with non- toxic
46、 metallic or plastic (screw) caps may be used. 6.7 Sterile Petri dishes, with diameters of approximately 55 mm and 90 mm. 6.8 Water bath, capable of operating at 47-50 C. 6.9 Water bath (or incubator), capable of operating at 50 C 2 C. 7 Sample It is important that the laboratory works with a pure c
47、ulture which has been biochemically confirmed as Salmonella spp. 8 Taxonomy of Salmonella 8.1 General Approximately every 7 years, the WHO Collaborating Centre for Reference and Research on Salmonella (Institut Pasteur, Paris) publishes an update of the “Antigenic formulae of the Salmonella serovars
48、”, which is the basis for assigning serovar names and formulas to isolates of Salmonella spp. At the time of publication, the latest version of the WhiteKauffmannLe Minor scheme is that of 2007 (Reference 9). NOTE Supplements to the White-Kauffmann-Le Minor scheme are published in Research in Microb
49、iology, a publication of the Institut Pasteur (formerly called Annales de lInstitut Pasteur/Microbiologie). For instance, supplement no. 47 was published in 2010 and characterises new serovars found between 2003 and 2007 (Reference 10). This part of ISO 6579 provides guidance on the serotyping of Salmonella serovars. 8.2 Nomenclature Different nomenclatures have been used (or are still in use) for Salmonella strains: originally, Kauffmann (Reference 12) considered each Salmonella serovar as a sep
