DIN EN 14476-2015 Chemical disinfectants and antiseptics - Quantitative suspension test for the evaluation of virucidal activity in the medical area - Test method and requirements .pdf

1、December 2015 English price group 18No part of this translation may be reproduced without prior permission ofDIN Deutsches Institut fr Normung e. V., Berlin. Beuth Verlag GmbH, 10772 Berlin, Germany,has the exclusive right of sale for German Standards (DIN-Normen).ICS 11.080.20!%IV“2385127www.din.de

2、DIN EN 14476Chemical disinfectants and antiseptics Quantitative suspension test for the evaluation of virucidal activity in the medical area Test method and requirements (Phase 2/Step 1);English version EN 14476:2013+A1:2015,English translation of DIN EN 14476:2015-12Chemische Desinfektionsmittel un

3、d Antiseptika Quantitativer Suspensionsversuch zur Bestimmung der viruziden Wirkung im humanmedizinischen Bereich Prfverfahren und Anforderungen (Phase 2, Stufe 1);Englische Fassung EN 14476:2013+A1:2015,Englische bersetzung von DIN EN 14476:2015-12Antiseptiques et dsinfectants chimiques Essai quant

4、itatif de suspension pour lvaluation de lactivit virucide dans le domaine mdical Mthode dessai et prescriptions (Phase 2/tape 1);Version anglaise EN 14476:2013+A1:2015,Traduction anglaise de DIN EN 14476:2015-12SupersedesDIN EN 14476:2013-10www.beuth.deDTranslation by DIN-Sprachendienst.In case of d

5、oubt, the German-language original shall be considered authoritative.Document comprises 42 pages 11.15 DIN EN 14476:2015-12 2 A comma is used as the decimal marker. National foreword This document (EN 14476:2013+A1:2015) has been prepared by Technical Committee CEN/TC 216 “Chemical disinfectants and

6、 antiseptics” (Secretariat: AFNOR, France), Working Group WG 1 “Human medicine” (Secretariat: DIN, Germany). The responsible German body involved in its preparation was DIN-Normenausschuss Medizin (DIN Standards Committee Medical), Working Committee NA 063-04-07 AA Chemische Desinfektionsmittel und

7、Antiseptika in der Humanmedizin. Amendments This standard differs from DIN EN 14476:2013-10 as follows: a) for the hygienic handrub and handwash method, a test for virucidal activity against enveloped viruses with Vacciniavirus has been added; b) the relationship between this European Standard and t

8、he EU Directive 93/42/EEC on medical devices (MDD) was added (Foreword and Annex ZA); c) the value of vnin C.1 has been corrected (0,001 instead of 0,000 1); d) the translation has been partially revised. Previous editions DIN EN 14476: 2005-08, 2007-02, 2013-10 EUROPEAN STANDARD NORME EUROPENNE EUR

9、OPISCHE NORM EN 14476:2013+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 (Phase2/Step 1) Antiseptiques et dsinfe

10、ctants chimiques - 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

11、 Bereich - Prfverfahren 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 Eu

12、ropean Standard 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 versi

13、ons (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,

14、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, Sw

15、itzerland, Turkey 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 nati

16、onal Members. Ref. 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

17、.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 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 asse

18、ssing 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 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

19、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 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

20、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 their presence in the human body in case of virus infection . 24 Annex B (informative) Detoxification of test mixtures by molecular sieving . 26

21、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 HR . 28 Annex C (informative) Calculation of the viral infectivity titre 29 C.1 Quantal tests Example of TCID50 determination by the Spearman-

22、Krber method . 29 DIN EN 14476:2015-12 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 .

23、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 example . 33 Annex D (informative) Presentation of test results of one active concentration . 35 Annex E (informative) Quantita

24、tive determination of formaldehyde concentrations . 38 Annex ZA (informative) !Relationship between this European Standard and the Essential Requirements of EU Directive 93/42/EEC“ 39 Bibliography . 40 DIN EN 14476:2015-12 EN 14476:2013+A1:2015 (E) 4 European foreword This document (EN 14476:2013+A1

25、: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 given the status of a national standard, either by publication of an identical text or by endorsement, at the latest by Marc

26、h 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 be the subject of patent rights. CEN and/or CENELEC shall not be held responsible for identifying any or all such patent righ

27、ts. 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 is indicated in the text by tags !“. !This document has been prepared under a mandate given to CEN by the European Commissio

28、n 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 integral part of this document.“ The document was revised to adapt it to the latest state of science, to correct errors and

29、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 a list of significant technical changes since the last edition: The scope was ex

30、panded 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 can be chosen within limits) that have to be performed to pass the test. An add

31、itional 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 virucidal activity against enveloped viruses with Vacciniavirus was added. The relationship between this European Standard and

32、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. Other methods to evaluate the efficacy of chemical disinfectants and antiseptics for different applications in the medical

33、area are in preparation. DIN EN 14476:2015-12 EN 14476:2013+A1:2015 (E) 5 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, Den

34、mark, 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. DIN EN 14476:

35、2015-12EN 14476:2013+A1:2015 (E) 6 Introduction This European Standard specifies a suspension test for 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

36、 application of the product including contact time, temperature, test organisms and interfering substances, 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 e

37、xperimental conditions. DINEN 14476:2015-12EN 14476:2013+A1:2015(E)7 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 homogeneous physically stable preparation when diluted with har

38、d 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 produced by adding the test organisms and interfering subs

39、tance. 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 and textile disinfection. This European Standard applies t

40、o 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 nursing homes; and may occur in the workplace and in the h

41、ome. 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 conditions in which they are used. NOTE 2 This method corresponds

42、 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 in this document and are indispensable for its applicatio

43、n. 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 used for the determination of bactericidal (including Leg

44、ionella), 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 definitions For the purposes of this document, the terms and

45、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 of infectious virus particles per unit volume (ml) DIN EN

46、14476:2015-12 EN 14476:2013+A1:2015(E)8 3.3 reference test for virus inactivation test 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 th

47、at induce a CPE (3.5) in 50 % of cell culture units 3.5 viral cytopathic effect CPE 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 multiplicat

48、ion of a single infectious virus particle 3.7 virus titre amount of infectious virus 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 Cla

49、use 5. DIN EN 14476:2015-12 EN 14476:2013+A1:2015(E)9 Table 1 Minimum and additional test conditions Test Conditions Hygienic handrub and handwash Instrument disinfection Surface disinfection Textile disinfection Minimum spectrum of test organisms Poliovirus Adenovirus Murine Norovirus Limited spectrum virucidal activity aAdenovirus Murine Norovirus !Virucidal activity against enveloped viruses bVacciniavirus“ Poliovirus Adenovirus Murine Norov