1、February 2009DEUTSCHE NORM English price group 16No part of this standard 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!$V(“1510592
2、www.din.deDDIN EN 14563Chemical disinfectants and antiseptics Quantitative carrier test for the evaluation of mycobactericidal ortuberculocidal activity of chemical disinfectants used for instruments inthe medical area Test method and requirements (phase 2, step 2)English version of DIN EN 14563:200
3、9-02Chemische Desinfektionsmittel und Antiseptika Quantitativer Keimtrgerversuch zur Prfung der mykobakteriziden oder tuberkulozidenWirkung chemischer Desinfektionsmittel fr Instrumente im humanmedizinischenBereich Prfverfahren und Anforderungen (Phase 2, Stufe 2)Englische Fassung DIN EN 14563:2009-
4、02www.beuth.deDocument comprises 38 pagesDIN EN 14563:2009-02 2 National foreword This standard has been prepared by Technical Committee CEN/TC 216 “Chemical disinfectants and antiseptics” (Secretariat: AFNOR, France), Working Group WG 1 “Human medicine” (Secretariat: DIN, Germany). The responsible
5、German body involved in its preparation was the Normenausschuss Medizin (Medical Standards Committee), Technical Committee NA 063-04-07 AA Chemische Desinfektionsmittel und Antiseptika in der Humanmedizin. EUROPEAN STANDARDNORME EUROPENNEEUROPISCHE NORMEN 14563November 2008ICS 11.080.20English Versi
6、onChemical disinfectants and antiseptics - Quantitative carrier testfor the evaluation of mycobactericidal or tuberculocidal activity ofchemical disinfectants used for instruments in the medical area -Test method and requirements (phase 2, step 2)Dsinfectants et antiseptiques chimiques - Essai quant
7、itatifde porte-germe pour lvaluation de lactivitmycobactricide ou tuberculocide des dsinfectantschimiques utiliss pour instruments en mdecine humaine -Mthode dessai et prescriptions (phase 2, tape 2)Chemische Desinfektionsmittel und Antiseptika -Quantitativer Keimtrgerversuch zur Prfung dermykobakte
8、riziden oder tuberkuloziden Wirkung chemischerDesinfektionsmittel fr Instrumente imhumanmedizinischen Bereich - Prfverfahren undAnforderungen (Phase 2, Stufe 2)This European Standard was approved by CEN on 18 October 2008.CEN members are bound to comply with the CEN/CENELEC Internal Regulations whic
9、h stipulate the conditions for giving this EuropeanStandard the status of a national standard without any alteration. Up-to-date lists and bibliographical references concerning such nationalstandards may be obtained on application to the CEN Management Centre or to any CEN member.This European Stand
10、ard exists in three official versions (English, French, German). A version in any other language made by translationunder the responsibility of a CEN member into its own language and notified to the CEN Management Centre has the same status as theofficial versions.CEN members are the national standa
11、rds bodies of Austria, Belgium, Bulgaria, Cyprus, Czech Republic, Denmark, Estonia, Finland,France, Germany, Greece, Hungary, Iceland, Ireland, Italy, Latvia, Lithuania, Luxembourg, Malta, Netherlands, Norway, Poland, Portugal,Romania, Slovakia, Slovenia, Spain, Sweden, Switzerland and United Kingdo
12、m.EUROPEAN COMMITTEE FOR STANDARDIZATIONCOMIT EUROPEN DE NORMALISATIONEUROPISCHES KOMITEE FR NORMUNGManagement Centre: rue de Stassart, 36 B-1050 Brussels 2008 CEN All rights of exploitation in any form and by any means reservedworldwide for CEN national Members.Ref. No. EN 14563:2008: EEN 14563:200
13、8 (E) 2 Contents Page Foreword3 Introduction .4 1 Scope 5 2 Normative references 5 3 Terms and definitions .5 4 Requirements.5 5 Test method6 5.1 Principle6 5.2 Materials and reagents6 5.3 Apparatus and glassware .9 5.4 Preparation of test organism suspensions and product test solutions 11 5.5 Proce
14、dure for assessing the mycobactericidal / tuberculocidal activity of the product .13 5.6 Experimental data and calculation.16 5.7 Verification of methodology .22 5.8 Expression of results and precision22 5.9 Interpretation of results conclusion .23 5.10 Test report 24 Annex A (informative) Reference
15、d strains in national collections.26 Annex B (informative) Examples of neutralizers of the residual antimicrobial activity of chemical disinfectants and antiseptics and rinsing liquids 27 Annex C (informative) Graphical representations of the test method .29 Annex D (informative) Example of a typica
16、l test report.32 Annex ZA (informative) Relationship between this European Standard and the Essential Requirements of EU Directive 93/42/EEC35 Bibliography 36 DIN EN 14563:2009-02 EN 14563:2008 (E) 3 Foreword This document (EN 14563:2008) has been prepared by Technical Committee CEN/TC 216 “Chemical
17、 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 May 2009, and conflicting national standards shall be withdrawn at the l
18、atest by May 2009. 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 has been prepared under a mandate given to CEN by the
19、 European Commission and the European Free Trade Association, and supports essential requirements of EC Directive(s). For relationship with EU Directive 93/42/EEC, see informative annex ZA, which is an integral part of this document. Other methods to evaluate the efficacy of chemical disinfectants a
20、nd antiseptics for different applications in the medical field are in preparation. A collaborative trial will be undertaken to provide a precision annex to this standard. According to the CEN/CENELEC Internal Regulations, the national standards organizations of the following countries are bound to i
21、mplement this European Standard: Austria, Belgium, Bulgaria, Cyprus, Czech Republic, Denmark, Estonia, Finland, France, Germany, Greece, Hungary, Iceland, Ireland, Italy, Latvia, Lithuania, Luxembourg, Malta, Netherlands, Norway, Poland, Portugal, Romania, Slovakia, Slovenia, Spain, Sweden, Switzerl
22、and and the United Kingdom. DIN EN 14563:2009-02 EN 14563:2008 (E) 4 Introduction This European Standard specifies a carrier test for establishing whether a chemical disinfectant for use on instruments (surgical instruments, anaesthesia material, endoscopes etc.) has a mycobactericidal or tuberculoc
23、idal activity in the area described in the scope. The laboratory test closely simulates practical conditions of application including pre-drying mycobacteria on a carrier, contact time, temperature, test organisms and interfering substances, i.e. conditions which may influence the action of chemical
24、 disinfectants in practical situations. The obligatory conditions are intended to cover general purposes and to allow reference between laboratories and product types. Each utilization concentration of the chemical disinfectant found by this test corresponds to defined experimental conditions. Howev
25、er, for some applications the recommendations of use of a product may differ and therefore additional test conditions need to be used. DIN EN 14563:2009-02 EN 14563:2008 (E) 5 1 Scope This European Standard specifies a test method and the minimum requirements for mycobactericidal or tuberculocidal a
26、ctivity of chemical disinfectant products that form a homogeneous, physically stable preparation when diluted with hard water, or in the case of ready-to-use products with water. This European Standard applies to products that are used in the medical area for disinfecting instruments by immersion ev
27、en if they are not covered by the EEC/93/42 Directive on Medical Devices. 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; i
28、n clinics of schools, kindergartens and 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. EN 14885 specifies in detail the relationship of the various tests to one another and to “u
29、se recommendations“. NOTE This method corresponds to a phase 2, step 2 test. 2 Normative references The following referenced documents are indispensable for the application of this document. For dated references, only the edition cited applies. For undated references, the latest edition of the refer
30、enced document (including any amendments) applies. EN 12353, Chemical disinfectants and antiseptics Preservation of test organisms used for the determination of bactericidal, mycobactericidal, sporicidal and fungicidal activity EN 14885, Chemical disinfectants and antiseptics Application of European
31、 Standards for chemical disinfectants and antiseptics 3 Terms and definitions For the purposes of this European Standard, the terms and definitions given in EN 14885 apply. 4 Requirements The product, when tested in accordance with Clause 5 under simulated clean conditions (0,3 g/l bovine albumin so
32、lution) or simulated dirty conditions (3,0 g/l bovine albumin solution, plus 3,0 ml/l washed sheep erythrocytes) according to its practical applications and under the obligatory test conditions, (one or two selected test organisms, 20 C, 60 min), shall demonstrate at least a decimal log (lg)reductio
33、n in counts of 4. The mycobactericidal activity shall be evaluated using the following two test organisms: Mycobacterium avium and Mycobacterium terrae. The tuberculocidal activity shall be evaluated using the following test organism: Mycobacterium terrae. DIN EN 14563:2009-02 EN 14563:2008 (E) 6 Wh
34、ere indicated, additional specific mycobactericidal or tuberculocidal activity shall be determined applying other contact times, temperatures and interfering substances in accordance with 5.5.1.1, in order to take into account intended specific use conditions. NOTE For these additional conditions, t
35、he concentration defined as a result can be lower than the one obtained under the obligatory test conditions. 5 Test method 5.1 Principle 5.1.1 A test suspension of mycobacteria in a solution of interfering substances is spread on a glass carrier. After drying the carrier is immersed into a sample o
36、f the product as delivered and/or diluted with hard water (for ready to use products: water). The carrier is maintained at 20 C 1 C for 60 min 10 s (obligatory test conditions). At the end of this contact time, the carrier is transferred into a neutralizer containing glass beads. The mycobacteria ar
37、e to be severed from the surface by shaking. The numbers of surviving mycobacteria in each sample are determined and the reduction is calculated. 5.1.2 The test is performed using Mycobacterium avium and Mycobacterium terrae or only Mycobacterium terrae as test organisms (obligatory test conditions)
38、. 5.1.3 Additional and optional contact times and temperatures are specified. Additional interfering substances may be used. 5.2 Materials and reagents 5.2.1 Test organisms The mycobactericidal activity shall be evaluated using the following two test-organisms1): Mycobacterium avium ATCC 15769 Mycob
39、acterium terrae ATCC 15755 The tuberculocidal activity shall be evaluated using only Mycobacterium terrae. NOTE See Annex A for strain reference in some other culture collections. 5.2.2 Culture media and reagents 5.2.2.1 General All weights of chemical substances given in this European Standard refe
40、r to the anhydrous salts. Hydrated forms may be used as an alternative, but the weights required shall be adjusted to allow for consequent molecular weight differences. The reagents shall be of analytical grade and/or appropriate for microbiological purposes. They shall be free from substances that
41、are toxic or inhibitory to the test organisms. 1) The ATCC numbers are the collection numbers of strains supplied by the American Type Culture Collections (ATCC). This information is given for the convenience of users of this European Standard and does not constitute an endorsement by CEN of the pro
42、duct named. DIN EN 14563:2009-02 EN 14563:2008 (E) 7 NOTE 1 To improve reproducibility, it is recommended that commercially available dehydrated material is used for the preparation of culture media. The manufacturers instructions relating to the preparation of these products should be rigorously fo
43、llowed. NOTE 2 For each culture medium and reagent a time limitation for use should be fixed. 5.2.2.2 Water The water shall be freshly glass distilled water and not demineralized water. If distilled water of adequate quality is not available, water for injections (see bibliographic reference 1) may
44、be used. Sterilize in the autoclave (5.3.1). Sterilization is not necessary if the water is used e.g. for preparation of culture media and subsequently sterilized. NOTE See 5.2.2.7 for the procedure to prepare hard water. 5.2.2.3 Middlebrook and Cohn 7H10 medium enriched by 10 % OADC (MCO) Middlebro
45、ok 7H10 agar powder 19,0 g Glycerol (C3H8O3) (see bibliographic reference 2) 5,0 ml Water (5.2.2.2) to 900,0 ml Heat to boiling to dissolve completely. Sterilize in the autoclave (5.3.1) and cool to 50 C to 55 C. Add 100 ml Middlebrook OADC enrichment under aseptic conditions. Fill 18 20 ml per plat
46、e (5.3.2.10). The pH of the medium shall be equivalent to 6,6 0,2 when measured at 25 C. NOTE In special circumstances (problems with neutralization see 5.5.1.2 and 5.5.1.3) it may be necessary to add neutralizer to MCO (see Annex B). It is not recommended to use neutralizer that causes opalescence
47、in the agar. 5.2.2.4 Diluent Tryptone Sodium Chloride Solution: Tryptone, pancreatic digest of casein 1,0 g Sodium chloride (NaCl) 8,5 g Water (5.2.2.2) to 1 000,0 mlSterilize in the autoclave (5.3.1). After sterilization the pH of the diluent shall be equivalent to 7,0 0,2 when measured at (20 1) C
48、. 5.2.2.5 Neutralizer The neutralizer shall be validated for the product being tested in accordance with 5.5.1 and 5.5.2. The neutralizer shall be sterile. NOTE Information on neutralizers that have been found to be suitable for some categories of products is given in Annex B. 5.2.2.6 Sterile defibr
49、inated sheep blood The sterile defibrinated sheep blood can be acquired from a commercial supplier or prepared according to EN 14820. DIN EN 14563:2009-02 EN 14563:2008 (E) 8 5.2.2.7 Hard water for dilution of products Prepare: Solution A: Dissolve 19,84 g anhydrous magnesium chloride (MgCl2) or an equivalent of hydrated magnesium chloride and 46,24 g anhydrous calcium chloride (CaCl2) or an equivalent of hydrated calcium chloride in water (5.2.2.2) and dilute to 1 000 ml. Sterilize in the au
