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BS EN 17272:2020+A1:2025 Chemical disinfectants and antiseptics — Methods of airborne room disinfection by automated process — Determination of bactericidal, mycobactericidal, sporicidal, fungicidal, yeasticidal, virucidal and phagocidal activities, 2026
- undefined
- European foreword
- Introduction
- 1 Scope
- 2 Normative references
- 3 Terms and definitions
- 4 Requirements [Go to Page]
- 4.1 Efficacy tests
- 4.2 Distribution tests
- 5 Test method [Go to Page]
- 5.1 Principle [Go to Page]
- 5.1.1 Preliminary test to validate absence of residual effect
- 5.1.2 Efficacy test
- 5.1.3 Distribution test
- 5.2 Materials and reagents [Go to Page]
- 5.2.1 Test organisms
- 5.2.2 Culture media and reagents [Go to Page]
- 5.2.2.1 General
- 5.2.2.2 Culture media for bacteria, mycobacteria, spores, fungal spores and yeasts [Go to Page]
- 5.2.2.2.1 Water
- 5.2.2.2.2 Agar for bacterial counts (Tryptone Soya Agar: TSA)
- 5.2.2.2.3 Reconstituted milk for use with sensitive test organisms
- 5.2.2.2.4 Agar for mycobacteria (refer to 5.2.1.7. and 5.2.1.8)
- 5.2.2.2.5 Medium for spores (refer to 5.2.1.9)
- 5.2.2.2.6 Malt extract agar (MEA)
- 5.2.2.2.7 Diluent for microbial suspensions [Go to Page]
- !Bacteria, yeast, fungi and spores"
- Mycobacteria: Tryptone salt !(5.2.2.2.7)" or distilled water (5.2.2.2.1).
- 5.2.2.2.8 Liquid for test organism recovery and rinsing liquid for membrane filtration
- 5.2.2.3 Culture media and reagents for preparation of virus (test organisms 5.2.1.12 to 5.2.1.14.)
- 5.2.2.4 Culture media and reagents for preparation of bacteriophages (test organisms 5.2.1.15 to 5.2.1.16.) [Go to Page]
- 5.2.2.4.1 M17 culture broth
- 5.2.2.4.2 M17 agar (underlay agar)
- 5.2.2.4.3 Overlay agar (surface agar, soft agar)
- 5.2.2.4.4 Bacteriophage diluent (based on Ringer's solution diluted to 1/4)
- 5.2.2.4.5 SM buffer
- 5.2.2.4.6 CaCl2 stock solutions (1 mol/l and 0,05 mol/l)
- 5.2.2.4.7 Lactic acid solution (10 % volume fraction)
- 5.2.2.4.8 Bacteriophage and virus recovery and rinsing liquid
- 5.2.2.5 Interfering substance (See Annex C)
- 5.2.3 Carriers [Go to Page]
- 5.2.3.1 General
- 5.2.3.2 Obligatory carriers
- 5.2.3.3 Additional carriers
- 5.3 Apparatus and glassware [Go to Page]
- 5.3.1 General
- 5.3.2 Usual microbiological laboratory equipment, and in particular, the following:
- 5.4 Preparation and counting of test suspensions [Go to Page]
- 5.4.1 Preparation of test suspensions (bacteria, spores, fungal spores, yeasts, mycobacteria) [Go to Page]
- 5.4.1.1 Preservation of test organisms and stock cultures
- 5.4.1.2 Working culture of test organisms [Go to Page]
- 5.4.1.2.1 General
- 5.4.1.2.2 Bacterial suspensions
- 5.4.1.2.3 Mycobacterial suspensions
- 5.4.1.2.4 Spore suspensions
- 5.4.1.2.5 Fungal spores and yeasts
- 5.4.1.3 Preparation of suspensions in interfering substance [Go to Page]
- 5.4.1.3.1 Bacteria (see 5.2.1.1 to 5.2.1.6)
- 5.4.1.3.2 !Mycobacterium avium and Mycobacterium terrae (refer to 5.2.1.7 and 5.2.1.8)
- 5.4.1.3.3 Bacterial spores (refer to 5.2.1.9)
- 5.4.1.3.4 Candida albicans (refer to 5.2.1.10)
- 5.4.1.3.5 Aspergillus brasiliensis (refer to 5.2.1.11)
- Figure 1 — View of conidiospores using an optical microscope — presence of smooth and spiny spores (unacceptable suspension)
- Figure 2 —View of conidiospores using an optical microscope — high concentration of characteristic mature spiny spores (acceptable suspension) [Go to Page]
- 5.4.1.4 Counting of suspensions prepared in interfering substance [Go to Page]
- 5.4.1.4.1 Bacteria
- 5.4.1.4.2 Mycobacteria
- 5.4.1.4.3 Bacterial spores
- 5.4.1.4.4 Fungal spores and yeasts
- 5.4.2 Preparation and counting of viral suspensions [Go to Page]
- 5.4.2.1 Viral stock suspension
- 5.4.2.2 Viral test suspension
- 5.4.2.3 Infectivity test [Go to Page]
- 5.4.2.3.1 Virus titre on suspended cells in microtitration plates
- 5.4.2.3.2 Virus titre on pre-established mono-cellular lawns in microtitration plates
- 5.4.2.3.3 Plaque assay method
- 5.4.3 Preparation and counting of bacteriophage test suspensions [Go to Page]
- 5.4.3.1 Host bacteria stock culture
- 5.4.3.2 Host bacteria working culture
- 5.4.3.3 Bacteriophage stock suspension
- 5.4.3.4 High-titre bacteriophage suspension
- 5.4.3.5 Preparation of bacteriophage suspension
- 5.4.3.6 Preparation of bacteriophages in interfering substance
- 5.5 Procedure for evaluating the automated airborne disinfection process activity [Go to Page]
- 5.5.1 Procedure for evaluating the process activity according to conditions of use furnished by the manufacturer (bacteria, mycobacteria, spores, fungal spores, yeasts, bacteriophages). [Go to Page]
- 5.5.1.1 Experimental conditions (obligatory, supplementary obligatory and additional)
- Table 1 — Manufacturers' intended use volume claims [Go to Page]
- 5.5.1.2 Survival controls of test organisms on carriers (efficacy and distribution tests) [Go to Page]
- 5.5.1.2.1 General
- 5.5.1.2.2 Contamination and drying of the control-carriers
- 5.5.1.2.3 Counting of the control-carriers
- 5.5.1.2.4 Bacteriophage counts on the control-carriers
- 5.5.1.3 Preliminary test to validate absence of residual effect (efficacy test and distribution tests) [Go to Page]
- 5.5.1.3.1 Recovering product residues on the carrier and testing for any potential antimicrobial effect
- 5.5.1.3.2 Screening for an inhibitory effect in the agar medium
- 5.5.1.3.3 Screening for an inhibitory effect in filter membranes
- 5.5.1.3.4 Screening for a carrier-related inhibitory effect in the agar medium
- 5.5.1.4 Efficacy and distribution tests [Go to Page]
- 5.5.1.4.1 Contamination of the test-carriers
- 5.5.1.4.2 Exposure of the test-carriers to the automated airborne disinfection process under test
- 5.5.1.4.3 Counting of bacteria, mycobacteria, bacterial spores, yeasts, fungal spores, and bacteriophages recovered
- 5.5.1.4.4 Incubation and counting of the test mixture
- 5.5.2 Virucidal activity tests [Go to Page]
- 5.5.2.1 General
- 5.5.2.2 Preliminary test to validate absence of residual effect [Go to Page]
- 5.5.2.2.1 Recovering product residues on the carrier and testing for any potential virucidal effect
- 5.5.2.2.2 !Cytotoxicity
- 5.5.2.2.3 Susceptibility of cells to viruses
- 5.5.2.2.4 Evaluating inhibition of product activity
- 5.5.2.3 Efficacy test [Go to Page]
- 5.5.2.3.1 Contamination of the carriers
- 5.5.2.3.2 Exposure of the test-carriers to automated airborne disinfection process
- 5.5.2.3.3 Determination of viral titre after exposure
- 5.5.2.4 Calculation and expression of results
- 5.5.3 Procedure for evaluating the mycobactericidal activity [Go to Page]
- 5.5.3.1 General
- 5.5.3.2 Contamination of the carriers
- 5.5.3.3 Exposure of the test-carriers to the product under test
- 5.5.3.4 Counts on the carriers
- 5.6 !Bacteria, mycobacteria, bacterial spores, yeasts, fungal spores and bacteriophages" [Go to Page]
- 5.6.1 General
- 5.6.2 Colony counting and limit thresholds
- 5.6.3 Determining number of test organisms N in bacterial, mycobacterial, bacterial spores, yeasts, fungal spores and bacteriophage suspensions (refer to 5.4)
- 5.6.4 Determining number of microorganisms T on control-carriers (refer to 5.5.1.2)
- 5.6.5 Determining values obtained in the preliminary test (refer to 5.5.1.3)
- 5.6.6 Determining the number of test organisms and plaques on test-carriers post- exposure and the reduction R (refer to 5.5.1.4.3)
- 5.7 Interpretation of results
- 5.8 Test report
- Table 2 — RESULTS: example for an automated process
- Annex A (normative) Summary of test requirements
- A.1 Efficacy tests
- Table A.1 — Efficacy tests
- A.2 Distribution tests
- Table A.2 — Distribution tests
- Annex B (normative) Distance between diffusion device and test-carriers (efficacy test)
- Table B.1 — Values applied with a tolerance of ± 10 % for volumes between 30 m3 and 150 m3
- Annex C (normative) Interfering substance
- Table C.1 — Conditions for products
- Annex D (normative) Preparation of spore stock suspensions of Bacillus subtilis
- D.1 Material and reagents
- D.1.1 Tryptone Glucose Broth (TGB):
- D.1.2 Yeast extract Agar (MYA):
- D.1.3 Centrifuge, capable of 10 000 gN acceleration
- D.2 Preparation of Bacillus spore stock suspensions
- Annex E (normative) Graphical representation of the test procedure (efficacy test)
- Figure E.1 — Control-carriers
- Figure E.2 — Preliminary test
- Figure E.3 — Preliminary test – Recovery of product residues
- Figure E.4 — Preliminary test – Exposure and recovery
- Figure E.5 — Scheme of a test room !(distribution test)"
- Figure E.6 — Scheme 1 of an example carrier position !(distribution test)"
- Figure E.7 — Scheme 2 of an example carrier position (not to scale) !(distribution test)"
- Bibliography [Go to Page]
