BS EN 14476-2013 Chemical disinfectants and antiseptics Quantitative suspension test for the evaluation of virucidal activity in the medical area Test method and requirements (Phas.pdf

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1、BSI Standards PublicationBS EN 14476:2013Chemical disinfectants andantiseptics Quantitativesuspension test for theevaluation of virucidal activityin the medical area Testmethod and requirements(Phase 2/Step 1)Copyright European Committee for Standardization Provided by IHS under license with CENNot

2、for ResaleNo reproduction or networking permitted without license from IHS-,-,-BS EN 14476:2013 BRITISH STANDARDNational forewordThis British Standard is the UK implementation of EN 14476:2013. Itsupersedes BS EN 14476:2005 which is withdrawn.The UK participation in its preparation was entrusted to

3、TechnicalCommittee CH/216, Chemical disinfectants and antiseptics.A list of organizations represented on this committee can beobtained on request to its secretary.This publication does not purport to include all the necessaryprovisions of a contract. Users are responsible for its correctapplication.

4、 The British Standards Institution 2013. Published by BSI StandardsLimited 2013ISBN 978 0 580 76780 7ICS 11.080.20Compliance with a British Standard cannot confer immunity fromlegal obligations.This British Standard was published under the authority of theStandards Policy and Strategy Committee on 3

5、1 December 2013.Amendments issued since publicationDate Text affectedCopyright European Committee for Standardization Provided by IHS under license with CENNot for ResaleNo reproduction or networking permitted without license from IHS-,-,-BS EN 14476:2013EUROPEAN STANDARD NORME EUROPENNE EUROPISCHE

6、NORM EN 14476 August 2013 ICS 11.080.20 Supersedes EN 14476:2005+A1:2006English Version Chemical disinfectants and antiseptics - Quantitative suspension test for the evaluation of virucidal activity in the medical area - Test method and requirements (Phase 2/Step 1) Antiseptiques et dsinfectants chi

7、miques - Essai quantitatif de suspension pour lvaluation de lactivit virucide dans le domaine mdical - Mthode dessai et prescriptions (Phase 2/tape 1) Chemische Desinfektionsmittel und Antiseptika - Quantitativer Suspensionsversuch zur Bestimmung der viruziden Wirkung im humanmedizinischen Bereich -

8、 Prfverfahren und Anforderungen (Phase 2, Stufe 1) This European Standard was approved by CEN on 5 July 2013. 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 alter

9、ation. 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

10、 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, Estoni

11、a, 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 STANDA

12、RDIZATION COMIT EUROPEN DE NORMALISATION EUROPISCHES KOMITEE FR NORMUNG Management Centre: Avenue Marnix 17, B-1000 Brussels 2013 CEN All rights of exploitation in any form and by any means reserved worldwide for CEN national Members. Ref. No. EN 14476:2013: ECopyright European Committee for Standar

13、dization Provided by IHS under license with CENNot for ResaleNo reproduction or networking permitted without license from IHS-,-,-BS EN 14476:2013EN 14476:2013 (E) 2 Contents Page Foreword 4 Introduction .5 1 Scope 6 2 Normative references 6 3 Terms and definitions .6 4 Requirements .7 5 Test method

14、s 8 5.1 Principle 8 5.2 Materials and reagents, including cell cultures 9 5.2.1 Test organisms 9 5.2.2 Culture media, reagents and cell cultures 9 5.3 Apparatus and glassware 12 5.4 Preparation of test organism suspensions and product test solutions . 14 5.4.1 Test organisms suspensions (test virus

15、suspension) 14 5.4.2 Product test solutions 14 5.5 Procedure for assessing the virucidal activity of the product . 15 5.5.1 General . 15 5.5.2 Test procedure 16 5.5.3 Modified method for ready-to-use products 17 5.5.4 Cytotoxicity caused by product test solutions 17 5.5.5 Control of efficiency of su

16、ppression of products activity . 18 5.5.6 Reference test for virus inactivation. 18 5.5.7 Titration of the virus control 19 5.5.8 Titration of test samples 19 5.6 Experimental data and calculation 19 5.6.1 Protocol of results 19 5.6.2 Calculation of infectivity titer (TCID50 or PFU) . 19 5.7 Verific

17、ation of the methodology 19 5.8 Expression of results . 20 5.8.1 General . 20 5.8.2 Calculation of the virucidal activity of products . 20 5.9 Test report . 20 Annex A (informative) Examples of viruses sorted according to their presence in the human body in case of virus infection 22 Annex B (inform

18、ative) Detoxification of test mixtures by molecular sieving. 24 B.1 Molecular sieving with SephadexTMLH 20 . 24 B.1.1 Principle . 24 B.1.2 Sephadex suspension 24 B.1.3 Procedure 24 B.2 Molecular sieving using MicroSpinTMS 400 HR 26 Annex C (informative) Calculation of the viral infectivity titre . 2

19、7 C.1 Quantal tests Example of TCID50determination by the Spearman-Krber method . 27 C.2 Plaque test . 28 C.3 Biometrical evaluation of experimental approaches and assessment of the disinfecting effect on the virus (reduction R): 28 C.3.1 General . 28 C.3.2 Calculating the virus titre with 95 % conf

20、idence interval . 29 C.3.3 Calculating the reduction and its 95 % confidence interval . 29 C.3.4 Calculating the average reduction (R(mi) and its 95 % confidence interval. . 30 C.3.5 Practical example 31 Copyright European Committee for Standardization Provided by IHS under license with CENNot for R

21、esaleNo reproduction or networking permitted without license from IHS-,-,-BS EN 14476:2013EN 14476:2013 (E) 3 Annex D (informative) Presentation of test results of one active concentration . 33 Annex E (informative) Quantitative determination of formaldehyde concentrations 36 Bibliography 37 Copyrig

22、ht European Committee for Standardization Provided by IHS under license with CENNot for ResaleNo reproduction or networking permitted without license from IHS-,-,-BS EN 14476:2013EN 14476:2013 (E) 4 Foreword This document (EN 14476:2013) has been prepared by Technical Committee CEN/TC 216 “Chemical

23、disinfectants and antiseptics”, 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 February 2014 and conflicting national standards shall be withdrawn at th

24、e latest by February 2014. 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 14476:2005+A1:2006. The documen

25、t was revised to adapt it to the latest state of science, to correct errors and ambiguities, to harmonise the structure and wording with other existing tests of CEN/TC 216 or in preparation and to improve the readability of the standard and thereby make it more understandable. The following list is

26、a list of significant technical changes since the last edition: The scope was expanded for the following fields of application within the medical area, i.e. products for textile disinfection. “Obligatory test conditions” were replaced by “minimum test conditions” (test temperatures and contact times

27、 can be chosen within limits) that have to be performed to pass the test. An additional modified method is described to test ready-to-use products in a higher concentration than 80 %, i.e. 9 7%. Data obtained using the former version of EN 14476 may still be used. Other methods to evaluate the effic

28、acy of chemical disinfectants and antiseptics for different applications in the medical area are in preparation. According to the CEN/CENELEC Internal Regulations, the national standards organisations of the following countries are bound to implement this European Standard: Austria, Belgium, Bulgari

29、a, 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, Turke

30、y and the United Kingdom. Copyright European Committee for Standardization Provided by IHS under license with CENNot for ResaleNo reproduction or networking permitted without license from IHS-,-,-BS EN 14476:2013EN 14476:2013 (E) 5 Introduction This European Standard specifies a suspension test for

31、establishing whether a chemical disinfectant or an antiseptic has a virucidal activity in the area and fields described in the scope. This laboratory test takes into account practical conditions of application of the product including contact time, temperature, test organisms and interfering substan

32、ces, i.e. conditions which may influence its action in practical situations. Each utilisation concentration of the chemical disinfectant or antiseptic found by this test corresponds to the chosen experimental conditions. Copyright European Committee for Standardization Provided by IHS under license

33、with CENNot for ResaleNo reproduction or networking permitted without license from IHS-,-,-BS EN 14476:2013EN 14476:2013 (E) 6 1 Scope This European Standard specifies a test method and the minimum requirements for virucidal activity of chemical disinfectant and antiseptic products that form a homog

34、eneous physically stable preparation when diluted with hard water or in the case of ready-to-use products, i. e, products that are not diluted when applied, with water. Products can only be tested at a concentration of 80 % (97 %, with a modified method for special cases) as some dilution is always

35、produced by adding the test organisms and interfering substance. This European Standard applies to products that are used in the medical area in the fields of hygienic handrub, hygienic handwash, instrument disinfection by immersion, surface disinfection by wiping, spraying, flooding or other means

36、and textile disinfection. This European Standard applies to areas and situations where disinfection is medically indicated. Such indications occur in patient care, for example: in hospitals, in community medical facilities, and in dental institutions; in clinics of schools, of kindergartens, and of

37、nursing homes; and may occur in the workplace and in the home. It may also include services such as laundries and kitchens supplying products directly for the patients. NOTE 1 The method described is intended to determine the activity of commercial formulations or active substances under the conditi

38、ons in which they are used. NOTE 2 This method corresponds to a phase 2, step 1 test. NOTE 3 EN 14885 specifies in detail the relationship of the various tests to one another and to “use recommendations”. 2 Normative references The following documents, in whole or in part, are normatively referenced

39、 in this document and are indispensable for its application. For dated references, only the edition cited applies. For undated references, the latest edition of the referenced document (including any amendments) applies. EN 12353, Chemical disinfectants and antiseptics Preservation of test organisms

40、 used for the determination of bactericidal (including Legionella), mycobactericidal, sporicidal, fungicidal and virucidal (including bacteriophages) activity EN 14885, Chemical disinfectants and antiseptics Application of European Standards for chemical disinfectants and antiseptics 3 Terms and def

41、initions For the purposes of this document, the terms and definitions given in EN 14885 and the following apply. 3.1 cytotoxicity morphological alteration of cells and/or their destruction or their reduced sensitivity to virus multiplication caused by the product 3.2 plaque forming units PFU number

42、of infectious virus particles per unit volume (ml) Copyright European Committee for Standardization Provided by IHS under license with CENNot for ResaleNo reproduction or networking permitted without license from IHS-,-,-BS EN 14476:2013EN 14476:2013 (E) 7 3.3 reference test for virus inactivation t

43、est with a defined product (e.g. formaldehyde) in parallel with a product under test for the internal control of the test 3.4 TCID5050 % infecting dose of a virus suspension or that dilution of the virus suspension that induce a CPE (3.5) in 50 % of cell culture units 3.5 viral cytopathic effect CPE

44、 morphological alteration of cells and/or their destruction as a consequence of virus multiplication 3.6 viral plaque area of lysis formed in a cell monolayer under semisolid medium due to infection by and multiplication of a single infectious virus particle 3.7 virus titre amount of infectious viru

45、s per unit volume present in a cell culture lysate or in a solution 4 Requirements The product shall demonstrate at least a decimal log (lg) reduction of 4 in virus titre when tested in accordance with Table 1 and Clause 5. Copyright European Committee for Standardization Provided by IHS under licen

46、se with CENNot for ResaleNo reproduction or networking permitted without license from IHS-,-,-BS EN 14476:2013EN 14476:2013 (E) 8 Table 1 Minimum and additional test conditions Test Conditions Hygienic handrub and handwash Instrument disinfection Surface disinfection Textile disinfection Minimum spe

47、ctrum of test organisms Poliovirus Adenovirus Murine Norovirus Limited spectrum virucidal activity a Adenovirus Murine Norovirus Poliovirus Adenovirus Murine Norovirus when temperature is 40 C or higher: only Parvovirus Poliovirus Adenovirus Murine Norovirus Parvovirus additional Any relevant test o

48、rganism Test temperature according to the manufacturers recommendation, but at / between 20 C 20 C and 70 C 4 C and 30 C 30 C and 70 C Contact time according to the manufacturers recommendation but between but no longer than but no longer than but no longer than 30 s and 120 s 60 min 5 min or 60 min b20 min Interfering substance clean conditions 0,3 g/l bovine albumin solution (hygienic handrub) c0,3 g/l bovine albumin solution and/or 0,3 g/l bovine albumin solution and/or dirty conditions 3,0 g/l bovine albumin solution plus 3,0 ml/l erythrocytes (hygieni