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

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)BS EN 14476:2013+A1:2015BS EN 14476:2013+A1:2015 BRITISH STANDARDNational forewordThis Brit

2、ish Standard is the UK implementation of EN 14476:2013+A1:2015. It supersedes BS EN 14476:2013 which is withdrawn.The start and finish of text introduced or altered by amendment is indicated in the text by tags. Tags indicating changes to CEN text carry the number of the CEN amendment. For example,

3、text altered by CEN amendment A1 is indicated by .The UK participation in its preparation was entrusted to Technical Committee CH/216, Chemical disinfectants and antiseptics.A list of organizations represented on this committee can be obtained on request to its secretary.This publication does not pu

4、rport to include all the necessary provisions of a contract. Users are responsible for its correct application. The British Standards Institution 2015. Published by BSI Standards Limited 2015ISBN 978 0 580 89188 5ICS 11.080.20Compliance with a British Standard cannot confer immunity from legal oblig

5、ations.This British Standard was published under the authority of the Standards Policy and Strategy Committee on 31 December 2013.Amendments/corrigenda issued since publicationDate Text affected30 September 2015 Implementation of CEN amendment A1EUROPEAN STANDARD NORME EUROPENNE EUROPISCHE NORM EN 1

6、4476:2013+A1 September 2015 ICS 11.080.20 Supersedes EN 14476:2013English 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 chimiques

7、 - 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 - Prfve

8、rfahren und Anforderungen (Phase 2, Stufe 1) This European Standard was approved by CEN on 5 July 2013 and includes Amendment 1 approved by CEN on 27 July 2015. CEN members are bound to comply with the CEN/CENELEC Internal Regulations which stipulate the conditions for giving this European Standard

9、the status of a 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, Fr

10、ench, 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, Bulgari

11、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

12、y and United Kingdom. EUROPEAN COMMITTEE FOR STANDARDIZATION COMIT EUROPEN DE NORMALISATION EUROPISCHES KOMITEE FR NORMUNG CEN-CENELEC Management Centre: Avenue Marnix 17, B-1000 Brussels 2015 CEN All rights of exploitation in any form and by any means reserved worldwide for CEN national Members. Re

13、f. No. EN 14476:2013+A1:2015 EEN 14476:2013+A1:2015 (E) 2 Contents Page European foreword . 4 Introduction 6 1 Scope 7 2 Normative references 7 3 Terms and definitions . 7 4 Requirements . 8 5 Test methods . 10 5.1 Principle 10 5.2 Materials and reagents, including cell cultures 10 5.2.1 Test organi

14、sms 10 5.2.2 Culture media, reagents and cell cultures . 11 5.3 Apparatus and glassware 14 5.4 Preparation of test organism suspensions and product test solutions 15 5.4.1 Test organisms suspensions (test virus suspension) . 15 5.4.2 Product test solutions . 15 5.5 Procedure for assessing the viruci

15、dal activity of the product . 16 5.5.1 General . 16 5.5.2 Test procedure 17 5.5.3 Modified method for ready-to-use products 18 5.5.4 Cytotoxicity caused by product test solutions . 18 5.5.5 Control of efficiency of suppression of products activity . 20 5.5.6 Reference test for virus inactivation . 2

16、0 5.5.7 Titration of the virus control . 20 5.5.8 Titration of test samples 20 5.6 Experimental data and calculation 21 5.6.1 Protocol of results 21 5.6.2 Calculation of infectivity titer (TCID50or PFU) 21 5.7 Verification of the methodology . 21 5.8 Expression of results . 22 5.8.1 General . 22 5.8

17、.2 Calculation of the virucidal activity of products . 22 5.9 Test report 22 Annex A (informative) Examples of viruses sorted according to their presence in the human body in case of virus infection . 24 Annex B (informative) Detoxification of test mixtures by molecular sieving . 26 B.1 Molecular si

18、eving with SephadexTMLH 20 26 B.1.1 Principle 26 B.1.2 Sephadex suspension 26 B.1.3 Procedure 26 B.2 Molecular sieving using MicroSpinTMS 400 HR . 28 Annex C (informative) Calculation of the viral infectivity titre 29 C.1 Quantal tests Example of TCID50 determination by the Spearman-Krber method . 2

19、9 EN 14476:2013+A1:2015 (E) 3 C.2 Plaque test . 29 C.3 Biometrical evaluation of experimental approaches and assessment of the disinfecting effect on the virus (reduction R): 30 C.3.1 General . 30 C.3.2 Calculating the virus titre with 95 % confidence interval . 31 C.3.3 Calculating the reduction an

20、d its 95 % confidence interval . 31 C.3.4 Calculating the average reduction (R(mi) and its 95 % confidence interval. 32 C.3.5 Practical example . 33 Annex D (informative) Presentation of test results of one active concentration . 35 Annex E (informative) Quantitative determination of formaldehyde co

21、ncentrations . 38 Annex ZA (informative) !Relationship between this European Standard and the Essential Requirements of EU Directive 93/42/EEC“ 39 Bibliography . 40 BS EN 14476:2013+A1:2015EN 14476:2013+A1:2015 (E) 2 Contents Page European foreword . 4 Introduction 6 1 Scope 7 2 Normative references

22、 7 3 Terms and definitions . 7 4 Requirements . 8 5 Test methods . 10 5.1 Principle 10 5.2 Materials and reagents, including cell cultures 10 5.2.1 Test organisms 10 5.2.2 Culture media, reagents and cell cultures . 11 5.3 Apparatus and glassware 14 5.4 Preparation of test organism suspensions and p

23、roduct test solutions 15 5.4.1 Test organisms suspensions (test virus suspension) . 15 5.4.2 Product test solutions . 15 5.5 Procedure for assessing the virucidal activity of the product . 16 5.5.1 General . 16 5.5.2 Test procedure 17 5.5.3 Modified method for ready-to-use products 18 5.5.4 Cytotoxi

24、city caused by product test solutions . 18 5.5.5 Control of efficiency of suppression of products activity . 20 5.5.6 Reference test for virus inactivation . 20 5.5.7 Titration of the virus control . 20 5.5.8 Titration of test samples 20 5.6 Experimental data and calculation 21 5.6.1 Protocol of res

25、ults 21 5.6.2 Calculation of infectivity titer (TCID50or PFU) 21 5.7 Verification of the methodology . 21 5.8 Expression of results . 22 5.8.1 General . 22 5.8.2 Calculation of the virucidal activity of products . 22 5.9 Test report 22 Annex A (informative) Examples of viruses sorted according to th

26、eir presence in the human body in case of virus infection . 24 Annex B (informative) Detoxification of test mixtures by molecular sieving . 26 B.1 Molecular sieving with SephadexTMLH 20 26 B.1.1 Principle 26 B.1.2 Sephadex suspension 26 B.1.3 Procedure 26 B.2 Molecular sieving using MicroSpinTMS 400

27、 HR . 28 Annex C (informative) Calculation of the viral infectivity titre 29 C.1 Quantal tests Example of TCID50 determination by the Spearman-Krber method . 29 EN 14476:2013+A1:2015 (E) 3 C.2 Plaque test . 29 C.3 Biometrical evaluation of experimental approaches and assessment of the disinfecting e

28、ffect on the virus (reduction R): 30 C.3.1 General . 30 C.3.2 Calculating the virus titre with 95 % confidence interval . 31 C.3.3 Calculating the reduction and its 95 % confidence interval . 31 C.3.4 Calculating the average reduction (R(mi) and its 95 % confidence interval. 32 C.3.5 Practical examp

29、le . 33 Annex D (informative) Presentation of test results of one active concentration . 35 Annex E (informative) Quantitative determination of formaldehyde concentrations . 38 Annex ZA (informative) !Relationship between this European Standard and the Essential Requirements of EU Directive 93/42/EE

30、C“ 39 Bibliography . 40 BS EN 14476:2013+A1:2015EN 14476:2013+A1:2015 (E) 4 European foreword This document (EN 14476:2013+A1:2015) has been prepared by Technical Committee CEN/TC 216 “Chemical disinfectants and antiseptics”, the secretariat of which is held by AFNOR. This European Standard shall be

31、 given the status of a national standard, either by publication of an identical text or by endorsement, at the latest by March 2016 and conflicting national standards shall be withdrawn at the latest by March 2016. Attention is drawn to the possibility that some of the elements of this document may

32、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 14476:2013“. This document includes Amendment 1 approved by CEN on 2015-07-27. The start and finish of text introduced or altered by amendment

33、 is indicated in the text by tags !“. !This document has been prepared under a mandate given to CEN by the European Commission and the European Free Trade Association, and supports essential requirements of EU Directive(s). For relationship with EU Directive(s), see informative Annex ZA, which is an

34、 integral part of this document.“ The document 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 i

35、t more understandable. The following list is 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 con

36、ditions” (test temperatures and contact times 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%; ! For the hygienic handrub and handwash method a test for vi

37、rucidal activity against enveloped viruses with Vacciniavirus was added. The relationship between this European Standard and the MDD was added (Foreword and Annex ZA). The value of vnin C.1 was corrected (0,001 instead of 0,0001).“ Data obtained using the former version of EN 14476 may still be used

38、. Other methods to evaluate the efficacy of chemical disinfectants and antiseptics for different applications in the medical area are in preparation. EN 14476:2013+A1:2015 (E) 5 According to the CEN-CENELEC Internal Regulations, the national standards organizations of the following countries are bou

39、nd 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, Portuga

40、l, Romania, Slovakia, Slovenia, Spain, Sweden, Switzerland, Turkey and the United Kingdom. BS EN 14476:2013+A1:2015EN 14476:2013+A1:2015 (E) 4 European foreword This document (EN 14476:2013+A1:2015) has been prepared by Technical Committee CEN/TC 216 “Chemical disinfectants and antiseptics”, the sec

41、retariat 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 March 2016 and conflicting national standards shall be withdrawn at the latest by March 2016. Attention is drawn

42、 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 14476:2013“. This document includes Amendment 1 approved by CEN on 2015-07-

43、27. The start and finish of text introduced or altered by amendment is indicated in the text by tags !“. !This document has been prepared under a mandate given to CEN by the European Commission and the European Free Trade Association, and supports essential requirements of EU Directive(s). For relat

44、ionship with EU Directive(s), see informative Annex ZA, which is an integral part of this document.“ The document 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 preparatio

45、n and to improve the readability of the standard and thereby make it more understandable. The following list is 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 disinfecti

46、on. “Obligatory test conditions” were replaced by “minimum test conditions” (test temperatures and contact times 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.

47、 9 7%; ! For the hygienic handrub and handwash method a test for virucidal activity against enveloped viruses with Vacciniavirus was added. The relationship between this European Standard and the MDD was added (Foreword and Annex ZA). The value of vnin C.1 was corrected (0,001 instead of 0,0001).“ D

48、ata obtained using the former version of EN 14476 may still be used. Other methods to evaluate the efficacy of chemical disinfectants and antiseptics for different applications in the medical area are in preparation. EN 14476:2013+A1:2015 (E) 5 According to the CEN-CENELEC Internal Regulations, the

49、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, Slovakia, Slovenia, Spain, Sweden, Switzerland, Turkey and the United Kingdom. BS EN 14476:2013+A1:2015EN 14476:2013+A1:2015 (E) 6 Introduction This Europ