EN 16616:2015

2003-01.Slovenski inštitut za standardizacijo. Razmnoževanje celote ali delov tega standarda ni dovoljeno.Chemisches Desinfektionsmittel und Antiseptika - Chemothermische Wäschedesinfektion - Prüfverfahren und Anforderungen (Phase 2, Stufe 2)Désinfectants chimiques et antiseptiques - Désinfection thermochimique du textile - Méthode d'essai et prescriptions (phase 2, étape 2)Chemical disinfectants and antiseptics - Chemical-thermal textile disinfection - Test method and requirements (phase 2, step 2)71.100.35Kemikalije za dezinfekcijo v industriji in domaChemicals for industrial and domestic disinfection purposes11.080.20Dezinfektanti in antiseptikiDisinfectants and antisepticsICS:Ta slovenski standard je istoveten z:EN 16616:2015SIST EN 16616:2015en,fr,de01-oktober-2015SIST EN 16616:2015SLOVENSKI
STANDARD
EUROPEAN STANDARD NORME EUROPÉENNE EUROPÄISCHE NORM
EN 16616
August 2015 ICS 11.080.20 English Version
Chemical disinfectants and antiseptics - Chemical-thermal textile disinfection - Test method and requirements (phase 2, step 2)
Désinfectants chimiques et antiseptiques - Désinfection thermochimique du textile - Méthode d'essai et prescriptions (phase 2, étape 2)
Chemisches Desinfektionsmittel und Antiseptika - Chemothermische Wäschedesinfektion - Prüfverfahren und Anforderungen (Phase 2, Stufe 2) This European Standard was approved by CEN on 3 July 2015.
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 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, 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, 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 United Kingdom.
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Referenced strains in national collections . 30 Annex B (informative)
Suitable neutralizers and rinsing liquids . 32 B.1 General . 32 B.2 Neutralizers . 32 B.3 Neutralizer added to the agar for counting . 33 Annex C (informative)
Graphical representations of the test method . 34 Annex ZA (informative)
Relationship between this European Standard and the Essential Requirements of EU Directive 93/42/EEC concerning medical devices . 35 Bibliography . 36
Preservation of test organisms used for the determination of bactericidal (including Legionella), mycobactericidal, sporicidal, fungicidal and virucidal (including bacteriophages) activity EN 13624, Chemical disinfectants and antiseptics
Quantitative suspension test for the evaluation of fungicidal or yeasticidal activity in the medical area
Test method and requirements (phase 2, step 1) EN 13727, bactericidal
EN 14348, Chemical disinfectants and antiseptics
Quantitative suspension test for the evaluation of mycobactericidal activity of chemical disinfectants in the medical area including instrument disinfectants
Test methods and requirements (phase 2, step 1) 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 definitions given in EN 14885 and the following apply. SIST EN 16616:2015

≥ 60°C and a defined liquor ratio) a reduction of Enterococcus faecium in and on germ carriers of more than
7 lg-units will be achieved. Further, no test organisms are to be detected in 100 ml washing/disinfection liquid. This includes fungicidal, mycobactericidal / tuberculocidal activity. 5 Test methods 5.1 Principle Germ carriers made of cotton fabric are contaminated with a test suspension of microorganisms in defibrinated sheep blood. After drying the carriers are transferred into cotton bags and then the disinfection process in the washing machine is performed at test temperatures either t < 60°C or ≥ 60°C. The process refers to the disinfection step without prewash. At the end of the disinfection step of the procedure, the bags with the carriers have to be taken out (see 5.3.2.18). Each carrier is transferred into a separate tube containing neutralizer and glass-beads. The bacteria should be recovered from the carriers by shaking. The number of surviving bacteria in each sample is determined and the reduction rate is calculated. 5.2 Materials and reagents 5.2.1 Test organisms The bactericidal activity shall be evaluated using the following strains as test organisms1) — Pseudomonas aeruginosa ATCC 15442 — Escherichia coli (K12) ATCC 10538 — Staphylococcus aureus ATCC 6538 — Enterococcus hirae ATCC 10541 — Enterococcus faecium
ATCC 6057
The yeasticidal/fungicidal activity shall be evaluated using the following test organisms: — Candida albicans ATCC 10231 — Aspergillus brasiliensis (formerly Aspergillus Niger ATCC 16404)
ATCC 16404
The mycobactericidal/tuberculocidal activity shall be evaluated using the following test organisms: — Mycobacterium avium ATCC 15769 — Mycobacterium terrae
ATCC 15755
NOTE See Annex A for strain reference in some other culture collections.
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 product named. SIST EN 16616:2015

Sterilize in the autoclave [5.3.2.1a)]. After sterilization the pH of the medium shall be equivalent to 7,2 ± 0,2 when measured at (20 ± 1) °C. In case of encountering problems with neutralization (5.5.1.2 and 5.5.1.3) it may be necessary to add neutralizer to the TSA. Annex B gives guidance on the neutralizers that may be used. 5.2.2.5 Tryptone Soy Broth (TSB) Tryptone, pancreatic digest of casein 15,0 g Soy peptone, papaic digest of soybean meal 5,0 g Sodium chloride (NaCl) 5,0 g Water (5.2.2.2) to 1 000,0 ml
Sterilize in the autoclave [5.3.2.1a)]. After sterilization the pH of the medium shall be equivalent to 7,2 ± 0,2 when measured at (20 ± 1) °C. 5.2.2.6 Brain Heart Infusion Agar (BHI) Example: Brain heart infusion 12,5 g Beef heart infusion 5,0 g Proteose-Peptone 10,0 g Glucose 2,0 g Sodium chloride (NaCl) 5,0 g Dinatriumhydrogenphosphate 2,5 g Agar 10,0 g Water (5.2.2.2) to 1 000,0 ml
Sterilize in the autoclave [5.3.2.1a)]. After sterilization the pH of the medium shall be equivalent to 7,2 ± 0,2 when measured at (20 ± 1) °C. SIST EN 16616:2015

Sterilize in the autoclave [5.3.2.1a)]. After sterilization the pH of the medium shall be equivalent to 5,6 ± 0,2 when measured at (20 ± 1) °C. 5.2.2.8 Middlebrook and Cohn 7H10 medium incl. 10 % OADC (7H10) Middlebrook 7H10 agar 19,0 g Glycerol 5,0 ml Water (5.2.2.2) To 900,0 ml
Heat to boiling to dissolve completely. Sterilize for 10 min in the autoclave [5.3.2.1a)] and cool to 50 °C to 55 °C. Add 100 ml Middlebrook OADC enrichment under aseptic conditions. The final pH of the medium shall be equivalent 6,6 ± 0,2 when measured at (25 ± 1) °C. 5.2.2.9 Diluent Tryptone, pancreatic digest of casein 1,0 g Sodium chloride (NaCl) 8,5 ml Water (5.2.2.2) to 1 000,0 ml
Sterilize in the autoclave [5.3.2.1a)]. After sterilization, the pH of the diluent shall be equivalent to 7,0 ± 0,2 when measured at (20 ± 1) °C. 5.2.2.10 Neutralizer The neutralizer shall be validated for the product being tested in accordance with 5.5.1.2, 5.5.1.3 and 5.5.2. It shall be sterile. NOTE Information on neutralizers that have been found to be suitable for some categories of products is given in Annex B. 5.2.2.11 Sterile defibrinated sheep blood The sterile defibrinated sheep blood can be acquired from a commercial supplier. 5.3 Apparatus and glassware 5.3.1 General Sterilize all glassware and parts of the apparatus that will come into contact with the culture media and reagents or the sample, except those which are supplied sterile, by one of the following methods: a) by moist heat, in the autoclave [5.3.2.1a)]; b) by dry heat, in the hot air oven [5.3.2.1b)]. SIST EN 16616:2015

2) Disposable sterile equipment is an acceptable alternative to reusable glassware. 3) Vortex® in an example of a suitable product available commercially. This information is given for the convenience of users of this European Standard and does not constitute an endorsement by CEN of this product. SIST EN 16616:2015

270 threads/dm each
The cotton control cloth shall be bleached, unfinished and not brightened. After three washes the maximum tensile strength, wet, in the warp should be (63 ± 5) daN. NOTE 2 Cotton proofed in accordance with DIN 53919 [6] fulfils the requirements. 5.3.2.17 Machine ballast load and preparation of ballast load Textile of poly cotton (65 % polyester / 35 % cotton) shall be used as ballast load. For special purposes textile of cotton (100 %) can be used. NOTE 1 The bounded liquor is higher in pure cotton than in poly cotton. Amount of ballast load: 70 % ± 10 % of the max. capacity (in kg). The ballast load should be washed for 30 min at 80 °C to 90 °C without additives following at least 3 rinsing steps and sterilized in the autoclave prior to first use. The ballast load should be used for not more than 100 washing cycles (incl. preparation and disinfection cycles). After disinfection testing the ballast load should be washed for 30 min at 80 °C to 90 °C following at least 5 rinsing steps (at least 5 min each) and sterilized in the autoclave prior to use. NOTE 2 If foam is present in last rinsing step additional rinsing might be necessary. 5.3.2.18 Washing machine and preparation of the machine The washing machine and additional equipment shall guaranty the following specifications: SIST EN 16616:2015

4) cfu/ml = colony forming unit(s) per millilitre
Figure 1 —Observation of conidiospores under light microscope: presence of smooth (a) and spiny (b) spores [inappropriate (not usable) suspension]
Figure 2 —Observation of conidiospores under light microscope: High concentration of characteristic mature spores with spiny aspect (appropriate suspension) If there are less than 75 % spiny conidiospores it may be due to the Aspergillus strain used or to the media. Therefore use another strain, preferably from another supplier, and/or try out another media; 2) the absence of conidiospore germination (check at least 10 fields of view); 3) if germinated conidiospores are present, discard the suspension; 4) the absence of mycelia fragments (check at least 10 fields of view). If mycelia are present, proceed to a second fritted filtration. If mycelia are still present, discard the suspension. c) Adjust the number of conidiospores in the suspension to 1,5 × 107 cfu/ml to 5,0 × 107 cfu/ml using the diluent (5.2.2.9), estimating the number of cfu by any suitable means. Use the suspension within 4 h in a water bath controlled at (20 ± 1) °C (5.3.2.2). In any case, adjust the temperature according to 5.5.1.4 only immediately before the start of the test (5.5.2). The use of a cell counting device for adjusting the number of cells is highly recommended. When using a suitable counting chamber, follow the instructions explicitly. Each laboratory should therefore produce calibration data to establish the relationship between the counts obtained using the counting device and the counts (5.5.2.9) obtained by the pour plate or the spread plate technique [d)]. Experienced laboratories found a better fit to the required number of conidiospores when the conidiospore suspension count in the device was 10 % to 50 % higher than the number aimed at. d) To get the test suspension at the appropriate cfu/ml, centrifuge it again and resuspend it with defibrinated sheep blood (5.2.2.11) in the same volume. SIST EN 16616:2015
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