Chemical disinfectants and antiseptics - Methods of airborne room disinfection by automated process - Determination of bactericidal, mycobactericidal, sporicidal, fungicidal, yeasticidal, virucidal and phagocidal activities

The method described herein is designed to determine the disinfectant activity of processes used in hospital, medical, pharmaceutical and cosmetics, veterinary, industrial and food processing areas.
The product trialled is designed to be diffused as gaseous molecules or solid or liquid-form dispersants.
NOTE Concerning automatic disinfectant processes:
the limits to use, especially in terms of ability to diffuse throughout the room (min and max effective volumes), shall be specified and stated in the test report;
certain automatic disinfection processes are only suitable for use in large-volume spaces well in excess of 150 m3.
Under these conditions, the systems cannot be tested in indoor spaces less than 150 m3 due to the high power of the jet spray; "additional experimental conditions" (see 5.5.1.1.f) should therefore be used and the tests should only be performed in test rooms with volumes different to the obligatory conditions.

Chemische Desinfektionsmittel und Antiseptika - Verfahren zur luftübertragenen Raumdesinfektion durch automatisierte Verfahren - Bestimmung der bakteriziden, mykobakteriziden, sporiziden, fungiziden, levuroziden, viruziden, tuberkuloziden, und Phagen-Wirksamkeit

Die beschriebenen Prüfverfahren sind zur Bestimmung der desinfizierenden Wirkung von verwendeten Prozessen 1) im humanmedizinischen Bereich, 2) im Veterinärbereich, 3) in den Bereichen Lebensmittel, Industrie, Haushalt und öffentliche Einrichtungen ausgelegt, bei denen automatisierte Verfahren zur Verteilung von Chemikalien durch Diffusion in der Luft ohne manuelles Auftragen des Desinfektionsmittel durch einen Anwender genutzt werden. Dieses Dokument umfasst die Desinfektion von porenfreien Oberflächen, aber nicht der Luft.
Das Ziel der beschriebenen Verfahren besteht im Desinfizieren der Oberflächen der gesamten Fläche, einschließlich der Außenflächen der in diesen Räumen befindlichen Ausrüstung. Speziell für die Desinfektion von Medizinprodukten ausgelegte Luftbehandlungen und Produkte oder Prozesse sind vom Anwendungsbereich dieses Dokuments ausgeschlossen. Die beschriebenen Prüfverfahren und Volumina stellen eine festgelegte Herausforderung dar.
Dieses Dokument gilt für Prozesse, für die die Wirkung bei den folgenden Gruppen von Mikroorganismen ausgewiesen wird:
-   vegetative Bakterien;
-   Mykobakterien;
-   bakterielle Sporen;
-   Hefen;
-   Schimmelpilzsporen;
-   Viren;
-   Bakteriophagen.
Dieses Dokument enthält keine Verfahren, bei denen die Wirkungsweise auf Eintauchen und/oder Zirkulieren, Fluten, Besprühen, Wischen oder anderen Vorgehensweisen, bei denen das Produkt direkt auf die Oberflächen aufgetragen und nicht durch die Luft verteilt wird, beruht.

Antiseptiques et désinfectants chimiques - Méthodes de désinfection des pièces par voie aérienne par des procédés automatisés - Détermination de l'activité bactéricide, fongicide, levuricide, sporicide, tuberculocide, mycobactéricide, virucide et phagocide

Les méthodes d’essai décrites sont destinées à déterminer l’activité désinfectante des procédés de désinfection de produits chimiques dans l’air, sans application manuelle du désinfectant par un opérateur, utilisés dans les secteurs 1) médical, 2) vétérinaire, 3) agro-alimentaire, industriel, domestique et collectivités. Le présent document couvre la désinfection de surfaces non poreuses mais pas celle de l’air.
Les procédés décrits ont pour objectif la désinfection des surfaces de l’ensemble de la zone traitée, y compris les surfaces externes des équipements contenus dans ces locaux. Le traitement de l’air et les produits ou procédés spécifiquement destinés à la désinfection des dispositifs médicaux sont exclus du domaine d’application du présent document. Les méthodes et volumes d’essai décrits fournissent un objectif défini.
Le présent document est applicable aux procédés déclarés actifs contre les groupes de micro-organismes suivants :
-   bactéries végétatives,
-   mycobactéries,
-   spores bactériennes,
-   levures,
-   spores fongiques,
-   virus,
-   bactériophages.
Le présent document ne couvre pas les procédés dont le mode d’action repose sur l’immersion et/ou la circulation, l’inondation, la pulvérisation, l’essuyage ou d’autres procédés dans lesquels le produit est directement appliqué sur les surfaces et non par dispersion dans l’air.

Kemična razkužila in antiseptiki - Metode za dezinfekcijo površin v prostorih z delci v zraku z avtomatiziranim postopkom - Določevanje baktericidne, mikobaktericidne, sporocidne, fungicidne, virucidne, tuberkulocidne in fagocidne aktivnosti ter aktivnosti kvasovk

General Information

Status
Published
Public Enquiry End Date
04-Oct-2018
Publication Date
25-May-2020
Current Stage
6060 - National Implementation/Publication (Adopted Project)
Start Date
22-May-2020
Due Date
27-Jul-2020
Completion Date
26-May-2020

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SLOVENSKI STANDARD
SIST EN 17272:2020
01-julij-2020
Kemična razkužila in antiseptiki - Metode za dezinfekcijo površin v prostorih z
delci v zraku z avtomatiziranim postopkom - Določevanje baktericidne,
mikobaktericidne, sporocidne, fungicidne, virucidne, tuberkulocidne in fagocidne
aktivnosti ter aktivnosti kvasovk

Chemical disinfectants and antiseptics - Methods of airborne room disinfection by

automated process - Determination of bactericidal, mycobactericidal, sporicidal,
fungicidal, yeasticidal, virucidal and phagocidal activities
Chemische Desinfektionsmittel und Antiseptika - Verfahren zur luftübertragenen
Raumdesinfektion durch automatisierte Verfahren - Bestimmung der bakteriziden,

mykobakteriziden, sporiziden, fungiziden, levuroziden, viruziden, tuberkuloziden, und

Phagen-Wirksamkeit

Antiseptiques et désinfectants chimiques - Méthodes de désinfection des pièces par voie

aérienne par des procédés automatisés - Détermination de l'activité bactéricide,

fongicide, levuricide, sporicide, tuberculocide, mycobactéricide, virucide et phagocide

Ta slovenski standard je istoveten z: EN 17272:2020
ICS:
11.080.20 Dezinfektanti in antiseptiki Disinfectants and antiseptics
71.100.35 Kemikalije za dezinfekcijo v Chemicals for industrial and
industriji in doma domestic disinfection
purposes
SIST EN 17272:2020 en,fr,de

2003-01.Slovenski inštitut za standardizacijo. Razmnoževanje celote ali delov tega standarda ni dovoljeno.

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SIST EN 17272:2020
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SIST EN 17272:2020
EN 17272
EUROPEAN STANDARD
NORME EUROPÉENNE
April 2020
EUROPÄISCHE NORM
ICS 11.080.20; 71.100.35
English Version
Chemical disinfectants and antiseptics - Methods of
airborne room disinfection by automated process -
Determination of bactericidal, mycobactericidal,
sporicidal, fungicidal, yeasticidal, virucidal and phagocidal
activities

Antiseptiques et désinfectants chimiques - Méthodes Chemische Desinfektionsmittel und Antiseptika -

de désinfection des pièces par voie aérienne par des Verfahren zur luftübertragenen Raumdesinfektion

procédés automatisés - Détermination de l'activité durch automatisierte Verfahren - Bestimmung der

bactéricide, fongicide, levuricide, sporicide, bakteriziden, mykobakteriziden, sporiziden,

tuberculocide, mycobactéricide, virucide et phagocide fungiziden, levuroziden, viruziden, tuberkuloziden, und

Phagen-Wirksamkeit
This European Standard was approved by CEN on 13 October 2019.

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, France, Germany, Greece, Hungary, Iceland, Ireland, Italy, Latvia, Lithuania, Luxembourg, Malta, Netherlands, Norway,

Poland, Portugal, Republic of North Macedonia, Romania, Serbia, Slovakia, Slovenia, Spain, Sweden, Switzerland, Turkey and

United Kingdom.
EUROPEAN COMMITTEE FOR STANDARDIZATION
COMITÉ EUROPÉEN DE NORMALISATION
EUROPÄISCHES KOMITEE FÜR NORMUNG
CEN-CENELEC Management Centre: Rue de la Science 23, B-1040 Brussels

© 2020 CEN All rights of exploitation in any form and by any means reserved Ref. No. EN 17272:2020 E

worldwide for CEN national Members.
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SIST EN 17272:2020
EN 17272:2020 (E)
Contents Page

European foreword ...................................................................................................................................................... 3

Introduction .................................................................................................................................................................... 4

1 Scope .................................................................................................................................................................... 5

2 Normative references .................................................................................................................................... 5

3 Terms and definitions ................................................................................................................................... 6

4 Requirements ................................................................................................................................................... 7

4.1 Efficacy tests ..................................................................................................................................................... 7

4.2 Distribution tests ............................................................................................................................................ 8

5 Test method ...................................................................................................................................................... 8

5.1 Principle ............................................................................................................................................................. 8

5.2 Materials and reagents ................................................................................................................................. 9

5.3 Apparatus and glassware ........................................................................................................................... 15

5.4 Preparation and counting of test suspensions ................................................................................... 18

5.5 Procedure for evaluating the automated airborne disinfection process activity ................. 29

5.6 Experimental data and calculations (bacteria, yeasts, fungal spores, bacterial spores,

bacteriophages and mycobacteria) ........................................................................................................ 40

5.7 Interpretation of results ............................................................................................................................. 44

5.8 Test report ....................................................................................................................................................... 45

Annex A (normative) Summary of test requirements .................................................................................... 49

Annex B (normative) Distance between diffusion device and test-carriers .......................................... 52

Annex C (normative) Interfering substance ...................................................................................................... 53

Annex D (normative) Preparation of spore stock suspensions of Bacillus subtilis ............................. 56

Annex E (normative) Graphical representation of the test procedure .................................................... 58

Bibliography ................................................................................................................................................................. 64

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SIST EN 17272:2020
EN 17272:2020 (E)
European foreword

This document (EN 17272:2020) has been prepared by Technical Committee CEN/TC 216 “Chemical

disinfectants and antiseptics”, the secretariat of which is held by AFNOR.

This document shall be given the status of a national standard, either by publication of an identical text

or by endorsement, at the latest by October 2020, and conflicting national standards shall be withdrawn

at the latest by October 2020.

Attention is drawn to the possibility that some of the elements of this document may be the subject of

patent rights. CEN shall not be held responsible for identifying any or all such patent rights.

This document describes a Phase 2 step 2 method designed:

— to check, under standardized laboratory conditions close to real-world practice, that the proposed

airborne surface disinfection processes meet the objective for which they were devised;

— to cross-compare different processes under reproducible conditions;

— to provide an experimental design within specified limits when real-world-practice conditions

depart from the conditions given in the text below.

According to the CEN-CENELEC Internal Regulations, the national standards organisations of the

following countries are bound to implement this European Standard: Austria, Belgium, Bulgaria,

Croatia, Cyprus, Czech Republic, Denmark, Estonia, Finland, France, Germany, Greece, Hungary, Iceland,

Ireland, Italy, Latvia, Lithuania, Luxembourg, Malta, Netherlands, Norway, Poland, Portugal, Republic of

North Macedonia, Romania, Serbia, Slovakia, Slovenia, Spain, Sweden, Switzerland, Turkey and the

United Kingdom.
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SIST EN 17272:2020
EN 17272:2020 (E)
Introduction

The purpose of this document is to describe a test method for assessing the disinfectant activity of

airborne surface disinfection processes under a specific experimental condition.
The proposed test method consists of 2 parts:

— Part 1 - Efficacy test: intended to ensure that minimum efficacy requirements are fulfilled for each

type of activity claimed and for the targeted application area(s) (CEN/TC 216 WG 1 and/or WG 2

and/or WG 3).

— Part 2 - Distribution test: intended to ensure efficacy of the process throughout the enclosure. It is

performed with a reference test organism at 4 sampling positions.

The processes concerned include those involving chemical disinfectants in dispersed gaseous, vapour

and/or aerosolised form.

Every automated airborne disinfection cycle/application is unique and the purpose of this document is

to provide a defined challenge for the automated airborne disinfection system to successfully meet in

order to be considered an efficacious process. This standard method should therefore be regarded as a

useful starting point and not as a validation for all intended treatments with a particular automated

airborne disinfection system.

The method is used to qualify the process, i.e. the device(s) and product(s) needed for implementation.

For such chemical processes, the combination of device and product cannot be separated.

For the defined test conditions the number of carriers and their test positions can be increased

according to specific needs of a given application or local requirements.
The manufacturer:
— specifies the limitations and precautions for use of the process;

— ensures that the specified test conditions are representative for the recommended application(s).

The aim of this document is to simulate practical conditions of airborne disinfection in a laboratory

situation; obligatory conditions are defined according to the test method defined below. Additional

conditions are also proposed.

The test report specifies and summarizes the conditions under which the tests are carried out.

Generally, the processes are implemented after a cleaning procedure and then tested, according to the

application areas, under clean or low-level soiling conditions. For specified applications and/or

according to the manufacturer recommendations, test methods with other interfering substance can

also be envisaged as additional conditions.

The tests described in this document are based on measuring the reduction (expressed as decimal

logarithm lg) in terms of numbers of surviving test organisms of different strains of bacteria,

mycobacteria, bacterial spores, fungal spores, yeasts, viruses or bacteriophages and under specified

conditions. Test organisms may be supplemented by other test organisms. The experimental design

described in this document is expected to be followed, but the conditions can be varied according to the

needs of the practical application(s).
This method can be used as a basis for biosecurity applications in laboratories.

CEN/TC 216 phase 2, step 1 suspension tests for evaluating the irreversible inactivation by the product

cannot be performed as the product is changed by the diffusion through the air (e.g. liquid state vs

vapour state).
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SIST EN 17272:2020
EN 17272:2020 (E)
1 Scope

The test methods described are designed to determine the disinfectant activity of processes used in the

1) medical area, 2) veterinary area, 3) food, industrial, domestic and institutional area using automated

processes for distributing chemicals by air diffusion with no operator manually applying the

disinfectant. This document covers the disinfection of nonporous surfaces but not that of the air.

The objective of the described processes is to disinfect the surfaces of the overall area including the

external surfaces of the equipment contained in such rooms. Air handling and products or processes

specifically designed for the disinfection of medical devices are excluded from the scope of this

document. The test methods and volumes described provide a defined challenge.

This document is applicable to processes for which activity is claimed against the following groups of

microorganisms:
— vegetative bacteria,
— mycobacteria,
— bacterial spores,
— yeasts,
— fungal spores,
— viruses,
— bacteriophages.

This document does not cover processes for which the mode of action is based on immersing and/or

circulation, flooding, spraying, wiping or other processes where the product is directly applied to the

surfaces and not via air dispersion.
2 Normative references

The following documents are referred to in the text in such a way that some or all of their content

constitutes requirements of this document. For dated references, only the edition cited applies. For

undated references, the latest edition of the referenced document (including any amendments) applies.

EN 10088-2, Stainless steels —Part 2: Technical delivery conditions for sheet/plate and strip of corrosion

resisting steels for general purposes

EN 12353, Chemical disinfectants and antiseptics — Preservation of test organisms 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
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SIST EN 17272:2020
EN 17272:2020 (E)
3 Terms and definitions

For the purposes of this document, the terms of EN 14885 and the following definitions apply:

ISO and IEC maintain terminological databases for use in standardization at the following addresses:

• IEC Electropedia: available at https://www.electropedia.org/
• ISO Online browsing platform: available at https://www.iso.org/obp
3.1
chemical process

process in which the active substance is a chemical agent (product) diffused in gas, liquid and/or solid

form

Note 1 to entry: The product and the diffusion system (device) cannot be evaluated separately.

3.2
automated airborne disinfection process

process diffusing a product in the form of a gas, vapour and/or an aerosol (excluding aqueous steam)

from a device, without the need for human intervention, targeting surfaces and not the air

3.3
airborne disinfection contact time
ADC time

time from the first release of the product (disinfectant) to the point where carriers are recovered or to

the point where aeration starts, if an aeration time is considered necessary

Note 1 to entry: The carriers can be recovered at the end of ADC time, or during the aeration time, with suitable

personal protective equipment (PPE) where necessary.
3.4
aeration time

period of time during which an air exchange of the enclosure achieves an appropriate concentration of

the product in the enclosure based on the manufacturer’s use instructions and risk assessment, to

enable the recovery of the carriers

Note 1 to entry: The duration of this aeration time is dependent of the air treatment system characteristics.

3.5
distribution test

placement of test-carriers loaded with test organisms in such a way that the distribution of a product by

the combination of a device (machine) and the product achieves its claimed activity throughout the

enclosure

Note 1 to entry: This test is performed after or in parallel with the efficacy test.

3.6
supplementary obligatory conditions

test conditions used instead of the obligatory conditions where the practical use of the process and the

manufacturers claims are clearly and unambiguously excluding the obligatory test conditions

3.7
sensitive test organism

test organism where the drying causes a lg reduction of more than 1,5 by the end of the aeration time

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SIST EN 17272:2020
EN 17272:2020 (E)
4 Requirements
4.1 Efficacy tests

The automated airborne disinfection process to be tested under the obligatory experimental conditions

defined in 5.5 shall lead to the following reductions in terms of numbers, expressed in decimal log (lg):

Bactericidal activity:

— 5 lg or greater reduction on test-carriers compared to control-carriers not exposed to the process,

for each of the four specified bacterial test organisms.
For medical area, refer to 5.2.1.2, 5.2.1.3, 5.2.1.4, 5.2.1.5.
For veterinary area, refer to 5.2.1.1, 5.2.1.2, 5.2.1.3, 5.2.1.6.

For food, industrial, domestic and institutional area, refer to 5.2.1.1, 5.2.1.2, 5.2.1.3, 5.2.1.4.

Mycobactericidal activity:

— 4 lg or greater reduction for medical area and food industry and laboratory area on test-carriers

comparative to control-carriers not exposed to the process for the two test organisms

implemented, refer to 5.2.1.15, 5.2.1.16.

— 4 lg or greater reduction for veterinary area on test-carriers comparative to control-carriers not

exposed to the process for the specified test organism, refer to 5.2.1.15.
Sporicidal activity:

— 4 lg or greater reduction for medical area, 3 lg for veterinary area and food, industrial, domestic and

institutional area on test-carriers compared to control-carriers not exposed to the process for the

specified bacterial spore test organism, refer to 5.2.1.7.
Fungicidal activity:

— 4 lg or greater for medical area, veterinary area and food, industrial, domestic and institutional area

on test-carriers compared to control-carriers not exposed to the process for the two specified

fungal test organisms (yeast and fungal spore), refer to 5.2.1.8 and 5.2.1.9.
Yeasticidal activity:

— 4 lg or greater reduction for medical area, veterinary area and food industry and laboratory area on

test-carriers comparative to control-carriers not exposed to the process for the specified yeast test

organism, refer to 5.2.1.8.
Virucidal activity:

— 4 lg or greater reduction for medical area and food industry and laboratory area on test-carriers

comparative to control-carriers not exposed to the process, for the two specified test organisms,

refer to 5.2.1.10 and 5.2.1.11.

— 4 lg or greater reduction for veterinary area on test-carriers comparative to control-carriers not

exposed to the process, for the specified test organism, refer to 5.2.1.12.
Phagocidal activity:

— 4 lg or greater reduction for food industry and laboratory area on test-carriers comparative to

control-carriers not exposed to the process for the specified two test organisms, refer to 5.2.1.13

and 5.2.1.14.
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SIST EN 17272:2020
EN 17272:2020 (E)
The requirements and test methods are summarized in Annex A.
The reductions are expressed as decimal logarithmic values (lg).

The reductions indicated in the table of Annex A are the minimum reductions to be obtained on test-

carriers in comparison to control-carriers not exposed to the process.

These activities may be determined independently, i.e. by separate testing of the different

microbiological groups, e.g. mycobacteria, and do not all shall be undertaken simultaneously. Each

activity can be claimed independently but passing both bactericidal and yeasticidal activity as described

in Annex A is the minimum requirement to claim compliance with this document.

These seven activities shall be determined under the standard-reference experimental conditions

defined in 5.5.

Additional specific bactericidal, mycobactericidal, sporicidal, fungicidal, yeasticidal, virucidal, and

phagocidal activity can, as appropriate, be determined under other conditions (e.g. other ADC times,

type of carriers, test organisms, volume of the enclosure, temperature, humidity), for specific intended

uses. These additional conditions shall be described, recorded and reported in the test report.

Even if an automated airborne disinfection system has passed all or part of the test method described

here, the system and its delivered cycles shall then be validated in the individual enclosure (e.g. hospital

room, or animal house) in practice to be disinfected using appropriate biological or validated chemical

indicators.
4.2 Distribution tests

The reduction in the number of viable bacterial cells throughout the enclosure shall be determined with

Staphylococcus aureus ATCC 6538 = CIP 4,83 (refer to 5.2.1.2) to achieve 5 lg or greater reduction on

each test–carrier. Test-carriers shall be located in accordance with Annex A, part 2. Test conditions shall

replicate those used in the efficacy tests.
5 Test method
5.1 Principle
5.1.1 Preliminary test to validate absence of residual effect

The aim of the preliminary test is to identify whether there is any residual activity due to residual

product transferred via the carriers into the subculture media (agar plates and/or membranes) and to

find a method for eliminating this effect. This procedure should ensure that the results of the efficacy

test are based only on the irreversible inactivation of the test organisms and not on an inhibitory

(static) effect.

Non-contaminated carriers are exposed to the process after deposition and drying of the interfering

substance used in the assay.

Recovery into 100 ml (20 ml for virucidal activity) of sterile liquid medium, and testing for

microbiostatic (e.g. bacteriostatic) effects due to traces of the product residing on the carriers, which

could generate an inhibitory effect in agar medium and/or on the filter membranes or a decrease in the

residual viral titre.
5.1.2 Efficacy test

Using test organism suspensions containing interfering substance, deposit 50 µl per carrier prepared as

described in 5.2.3.

Spread and air-dry the inoculum as described in 5.5.1.2.2, then expose the prepared test-carriers to the

product diffused by the tested automated airborne disinfection process, under defined conditions.

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EN 17272:2020 (E)

Recover the surviving bacteria, mycobacteria, bacterial spores, yeasts, fungal spores, viruses or

bacteriophages by mechanical action, such as scraping (e.g. with a glass pipette or scalpel), or if

required sonicate, from the carriers, into the recovery liquid:

For bacteria, mycobacteria, bacterial spores, fungal spores and yeasts, dilute and inoculate the agar with

a fraction of the recovery liquid. Incubate and count the colonies. Filtrate the remaining recovery liquid

using membrane filtration, then rinse to eliminate as much product as possible. Transfer onto agar

medium. Place the carrier into agar medium to capture any remaining surviving test organisms.

Incubate and count the number of colonies.

For viruses and bacteriophages, dilute a fraction of the recovery liquid, incubate on a cell line and

determine the viral titre.
5.1.3 Distribution test

The distribution test is a replication of the efficacy test except that only Staphylococcus aureus is used as

the test organism and the test carriers are located in other defined positions and orientations.

5.2 Materials and reagents
5.2.1 Test organisms

Depending on the type of activity targeted, tests shall use all or some of the following test organisms.

These test organisms can be obtained from culture collections. The test organisms are:

For bactericidal activity tests:
— 5.2.1.1 Pseudomonas aeruginosa ATCC 15442 = CIP 103-467 (DSM 937)
— 5.2.1.2 Staphylococcus aureus ATCC 6538 = CIP 4.83 (DSM 799)
— 5.2.1.3 Enterococcus hirae ATCC 10541 = CIP 5855 (DSM 3320)
— 5.2.1.4 Escherichia coli ATCC 10536 = CIP 54127 (DSM 682)
— 5.2.1.5 Acinetobacter baumanii ATCC 19606 = CIP 70.34 (DSM 30007)
— 5.2.1.6 Proteus hauseri ATCC 13315 = CIP 58.60(DSM 30118)
For sporicidal activity tests:
— 5.2.1.7 Bacillus subtilis spores ATCC 6633 = CIP 52 62
For yeasticidal activity tests:
— 5.2.1.8 Candida albicans ATCC 10231 = IP 4872 (DSM 1386)
For fungicidal activity tests:
— 5.2.1.8 Candida albicans ATCC 10231 = IP 4872 (DSM 1386)
— 5.2.1.9 Aspergillus brasiliensis ATCC 16404 = IP 1431-83 (DSM 1988)
For virucidal activity tests:
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SIST EN 17272:2020
EN 17272:2020 (E)

— 5.2.1.10 Murine Norovirus souche S99, Friedrich Loefler Institut, Berlin. MNV cultured on

RAW 264.7 (ATCC TIB-71) cells line

— 5.2.1.11 Adenovirus type 5, adenoid strain, ATCC VR-5. Adenovirus cultured on HeLa cells or other

lines of suitable susceptibility.

— 5.2.1.12 Porcine Parvovirus NADL2 strain cultured on ST cells or other appropriate cells line

For phagocidal activity:

— 5.2.1.13 Bacteriophage for Lactococcus lactis subspecies lactis P001 (DSM 4262)

— 5.2.1.14. Bacteriophage for Lactococcus lactis subspecies lactis P008 (DSM 10567)

The multiplication of these two bacteriophages shall be obtained from the following host strain:

Lactococcus lactis subspecies lactis F7/2 (DSM 4366).
For mycobactericidal activity:
— 5.2.1.15. Mycobacterium avium ATCC 15769 = CIP 105415
— 5.2.1.16 Mycobacterium terrae ATCC 15755 (tuberculocidal activity)

Both test organisms shall be tested, except for the veterinary area where only Mycobacterium avium is

required.
These strains can be obtained from biological collection resource centres.
The use of other test organisms

If additional test organisms are used, they shall be incubated under optimum growth conditions

(temperature, time, atmosphere, media) and noted in the test report. If the additional test organisms

selected do not correspond to the specified strains, their suitability for supplying the required inocula

shall be verified. If these additional test organisms are not classified at a reference centre, their

identification characteristics shall be stated. In addition, they shall be held by the testing laboratory or

national culture collection under a reference for five years.
5.2.2 Culture media and reagents
5.2.2.1 General

All weights of chemical substances given in this document refer 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 are toxic or inhibitory to the test organisms.

NOTE Commercial information (e.g. product name…) are provided by CEN as a convenience to users of this

document and does not represent an endorsement by CEN of this product. Equivalent products can be used if it is

demonstrated that they lead to the same results.

5.2.2.2 Culture media for bacteria, mycobacteria, spores, fungal spores and yeasts

5.2.2.2.1 Water
The water shall be freshly glass-distilled water and not demineralized water.
Sterilize in the autoclave (5.3.2.1a).
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SIST EN 17272:2020
EN 17272:2020 (E)

Sterile distilled water is necessary for the preparation of suspensions for tests. Sterilization is not

necessary if the water is used for e.g. preparation of culture media and subsequently sterilized.

NOTE If distilled water of adequate quality is not available, water for injection can be used.

5.2.2.2.2 Agar for bacterial counts (Tryptone Soya Agar: TSA)
For counts of viable bacterial cells (5.2.1.1 to 5.2.1.6)
Tryptone, pancreatic digest of casein 15,0 g
Soya peptone, papaic digest of soybean meal 5,0 g
NaCl 5,0 g
Agar 15,0 g
Water (see 5.2.2.2.1) to 1 000 ml

Sterilize in the autoclave. After sterilization, pH of the medium shall be equivalent to (7,2 ± 0,2) when

measured at (20 ± 1) °C.

If necessary, add a neutralizer to the medium (and record the addition in the test report).

Agar for the preservation of test organisms 5.2.1.1 to 5.2.1.6 (refer to 5.2.2.2.2) without neutralizer.

5.2.2.2.3 Reconstituted milk for use with sensitive test organisms
Prepare the reconstituted skimmed milk (15 g/l fat) as follows:

Skimmed powdered milk, guaranteed antibiotic-free and additive-free, reconstituted at 100 g powder

per 1 l distilled water.
Sterilize 30 min at (105 ± 3) °C or 5 min at (121 ± 3) °C.
5.2.2.2.4 Malt extract agar (MEA)

For counts on the number of viable yeast and fungal spore cells (5.2.1.8 and 5.2.1.9).

Malt extract (technical grade) 30,0 g
Agar 15,0 g
Water
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