ASTM E2721-2016 Standard Practice for Evaluation of Effectiveness of Decontamination Procedures for Surfaces When Challenged with Droplets Containing Human Pathogenic Viruses《当面对含有.pdf

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1、Designation: E2721 10E2721 16Standard Test Method Practice forEvaluation of Effectiveness of Decontamination Proceduresfor Surfaces When Challenged with Droplets ContainingHuman Pathogenic Viruses1This standard is issued under the fixed designation E2721; the number immediately following the designa

2、tion indicates the year oforiginal adoption or, in the case of revision, the year of last revision. A number in parentheses indicates the year of last reapproval. Asuperscript epsilon () indicates an editorial change since the last revision or reapproval.INTRODUCTIONMany communicable diseases can of

3、ten spread through droplets containing infectious agents. Such“contagious droplets” may expose susceptible individuals directly or contaminate environmentalsurfaces in the immediate vicinity and render them as fomites for further spread of the disease. Thecharacteristics of the droplets (particle si

4、ze and composition) will influence the viability of themicroorganisms when exposed to environmental stresses but also shield them from physical andchemical decontaminants. The wide variations in the types and levels of such protective/shieldingingredients can impact on the effectiveness of surface d

5、econtaminants. This test method practice isdesigned to simulate surface deposition of contagious droplets from human respiratory secretions. Itis primarily focused on influenza viruses but other respiratory viruses or surrogates could be used.Protocols for assessing the microbicidal activity of disi

6、nfectants are also described.1. Scope1.1 This test method is designed to evaluate decontamination methods (physical, chemical, self-decontaminating materials)when used on surfaces contaminated with virus-containing droplets.1.2 This test method defines the conditions for simulating respiratory dropl

7、ets produced by humans and depositing the dropletsonto surfaces.1.3 The method is specific to influenza viruses but could be adapted for work with other types of respiratory viruses orsurrogates (Appendix X5).1.4 This test method is suitable for working with a wide variety of environmental surfaces.

8、1.5 This test method does not address the performance of decontaminants against microbes expelled via blood splatter, vomit,or fecal contamination.1.6 This test method should be performed only by those trained in bioaerosols, microbiology, or virology, or combinationsthereof.1.7 The values stated in

9、 SI units are to be regarded as standard. No other units of measurement are included in this standard.1.8 This standard does not purport to address all of the safety concerns, if any, associated with its use. It is the responsibilityof the user of this standard to establish appropriate safety and he

10、alth practices and determine the applicability of regulatorylimitations prior to use.2. Referenced Documents2.1 ASTM Standards:2E1052 Test Method to Assess the Activity of Microbicides against Viruses in Suspension1 This test method practice is under the jurisdiction ofASTM Committee E35 on Pesticid

11、es,Antimicrobials, andAlternative ControlAgents and is the direct responsibilityof Subcommittee E35.15 on Antimicrobial Agents.Current edition approved Oct. 1, 2010April 1, 2016. Published February 2011May 2016. Originally approved in 2010. Last previous edition approved in 2010 as E272110.DOI: 10.1

12、520/E272110.10.1520/E272116.2 For referencedASTM standards, visit theASTM website, www.astm.org, or contactASTM Customer Service at serviceastm.org. For Annual Book of ASTM Standardsvolume information, refer to the standards Document Summary page on the ASTM website.This document is not an ASTM stan

13、dard and is intended only to provide the user of an ASTM standard an indication of what changes have been made to the previous version. Becauseit may not be technically possible to adequately depict all changes accurately, ASTM recommends that users consult prior editions as appropriate. In all case

14、s only the current versionof the standard as published by ASTM is to be considered the official document.Copyright ASTM International, 100 Barr Harbor Drive, PO Box C700, West Conshohocken, PA 19428-2959. United States1E2197 Quantitative Disk Carrier Test Method for Determining Bactericidal, Virucid

15、al, Fungicidal, Mycobactericidal, andSporicidal Activities of ChemicalsE2720 Practice for Evaluation of Effectiveness of Decontamination Procedures for Air-Permeable Materials when Challengedwith Biological Aerosols Containing Human Pathogenic Viruses2.2 EPA Standards:EPA 6004-84013 (N16) USEPA Manu

16、al of Methods for Virology32.3 WHO Standards:WHO Manual on Animal Influenza Diagnosis and Surveillance43. Terminology3.1 Definitions:3.1.1 aerosol, na suspension of solid or liquid particles in a gas medium.3.1.2 biological aerosol, naerosol comprising particles of biological origin or activity whic

17、h may affect living things throughinfectivity, allergencity, toxicity, or pharmacological and other processes.3.1.3 contact transmission, ninfections caused by direct skin-to-skin contact or indirect contact with objects contaminatedwith pathogens.3.1.4 contagious respiratory droplet, nrespiratory s

18、ecretions containing infectious microorganisms that form large droplets(5 m) and settle out of the air over short distances.3.1.5 droplet transmission, ndirect transfer of pathogen-containing droplets to conjuncitval or mucous membranes.3.1.6 influenza, nan infectious disease of birds and mammals ca

19、used by RNA viruses of the family Orthomyxoviridae.3.1.7 protective factor, nsoluble or insoluble material co-deposited with microorganisms that directly protects themicroorganism from environmental stresses or decontaminants.3.1.8 self-sanitizing material, na substrate containing an antimicrobial a

20、gent that collectively acts as a germicide.4. Summary of Test Method4.1 The test method describes the steps required to deposit droplets onto surfaces and quantitatively assess decontaminationefficiency.4.1.1 Using an aerosol device capable of meeting the data quality objectives set for in this test

21、 method, influenza virus orsurrogates are aerosolized to form droplets that are subsequently applied to surfaces.4.1.2 The virus-contaminated carriers are subjected to disinfection protocols and incubated for the specified time and conditions.Control samples are incubated under identical conditions

22、but are not exposed to the disinfection protocols.NOTE 1Carriers with incorporated microbicides do not receive any additional disinfection treatment. An untreated control is needed to assessantimicrobial efficacy.4.1.3 Virus particles are eluted from the test and control carriers and viability is as

23、sessed by 50 % tissue culture infectious doseassay (log10TCID50).NOTE 2Nonviable techniques for viral enumeration such as polymerase chain reaction (PCR) or hemagglutination cannot be used.4.1.4 The virucidal activity of the decontamination procedure is determined from the log difference in viabilit

24、y between treatedand test carriers.5. Significance and Use5.1 The efficacy of disinfection technologies can be evaluated on finished products, as well as on developmental items.5.2 This test method defines procedures for validation of the droplet generator, preparation of the test specimen, applicat

25、ion ofthe challenge virus, enumeration of viable viruses, assessing data quality, and calculation of decontamination efficiency.5.3 This test method provides defined procedures for creating droplets that approximate those produced by human respiratorysecretions, with particular emphasis on droplet s

26、ize distribution and aerosolization media.5.4 Safety concerns associated with aerosolizing microbial agents are not addressed as part of this test method. Individual usersshould consult with their local safety authority, and a detailed biological aerosol safety plan and risk assessment should becond

27、ucted prior to using this method. Users are encouraged to consult the manual Biosafety in Microbiological and BiomedicalLaboratories5 published by the U.S. Centers for Disease Control and Prevention (CDC).3 Available from United States Environmental Protection Agency (EPA), Ariel Rios Bldg., 1200 Pe

28、nnsylvania Ave., NW, Washington, DC 20460, http:/www.epa.gov.4 Webster, R., Cox, N., Stohr, K. WHO Manual on Animal Influenza Diagnosis and Surveillance. World Health Organization, Department of Communicable DiseaseSurveillance and Response. WHO/CDS/CDR/2002.5 Rev. 1.5 CDC-NIH, Biosafety in Microbio

29、logical and Biomedical Laboratories, 5th Edition, U.S. Department of Health and Human Services, Washington, D.C., 2009.E2721 1625.5 This test method differs from Test Methods E1052 and E2197 in the presentation of virus to the surface. The aforementionedtest methods use liquid inoculum to contaminat

30、e carrier surfaces, whereas this method presents the virus in droplets that arerepresentative of human respiratory secretions5.6 This method differs from Test Method E2720, because (1) larger droplets are being formed, (2) the droplets will not becompletely dried prior to application to surfaces, (3

31、) the droplets can be applied to any surfaces, not just those that are airpermeable, and (4) unique equipment is required to create droplets.6. Apparatus6.1 Droplet ApparatusThe apparatus used to load microorganisms onto a substrate is composed of several commerciallyavailable components and can be

32、readily constructed.6,7,8 The overall design of the apparatus can take various forms and can befashioned in different dimensions while meeting the validation requirements and data quality objectives listed below. Appendix X1contains the description of a prototypical device that can be used to load d

33、roplets onto surfaces. However, it is the responsibilityof the user of this standard to validate the performance of the device prior to use.6.1.1 Validation requirements and baseline testing.6.1.1.1 Environmental ConditionsGenerator must be capable of delivering air with a relative humidity of 50 6

34、10 %.6.1.1.2 Loading uniformity across the diameter of the test specimen is required to ensure the even distribution of the dropletsover the surface of the carrier. A standard deviation of 60.5 log10TCID50 is desired.6.1.1.3 Sample-to-Sample Variation ObjectiveThe variability of virus loading for mu

35、ltiple samples loaded for a single testmust have a standard deviation of 60.5 log10 TCID50.6.1.1.4 Droplet CharacteristicsThe droplets generated for this method will have a number median diameter (CMD) of 156 5 m. The virus will be aerosolized in a saliva substitute (Table 1) that will add the appro

36、priate “protective factors.” This methodwould be suitable for simulating other fluids of interest; however, if a different fluid is used, the formulation and recipe listing theprotective factors and droplet size must be reported.6.2 Other EquipmentThe list is specific for influenza virus. Other equi

37、pment may be needed if a different virus is used.6.2.1 Autoclave, capable of maintaining 121 to 123C and 15 to 17 lbs per in.2-gauge (psig).6.2.2 CO2 Incubator, capable of maintaining 35 to 37C and 5 6 0.5 % CO2.6.2.3 Vortex Mixer.6.2.4 Analytical Balance, capable of weighing 0.001 g.6.2.5 Refrigera

38、tor, capable of maintaining 2 to 8C.6.2.6 Stopwatch or Electronic Timer.6.2.7 Pipettor, with a precision of 0.001 mL.7. Reagents and Materials7.1 ReagentsThe list is specific for influenza use. Other reagents may be needed if a different virus is used.7.1.1 Influenzavirus (H1N1; A/PR/8/34)cell cultu

39、re adapted, ATCC VR-1469.6 Vo, E., Rengasamy, S., Shaffer, R., “Development of a Test System to Evaluate Decontamination Procedures for Viral Droplets on Respirators.” Applied andEnvironmental Microbiology, Vol 75, No. 23, 2009, pp. 73037309.7 Woo, M. H., Hsu,Y. M., Wu, C.Y., Heimbuch, B. K., Wander

40、, J. D., “ADevice for a Consistent and Controlled Delivery ofAerosolized Droplets Containing ViralAgentsOnto Surfaces.” Journal of Aerosol Science, Vol 41, 2010, pp. 941-952.8 Heimbuch B. K., Wallace, W. H., Kinney, K., Lumley,A. E., Wu, C-Y, Woo, M-H, Wander, J. D., “APandemic Influenza Preparednes

41、s Study: Use of Energetic Methodsto Decontaminate Filtering Facepiece Respirators Contaminated with H1N1 Aerosols and Droplets,” American Journal of Infection Control, 2010, DOI 10.1016/j.ajic.2010.07.004.TABLE 1 Artificial Saliva8Reagent AmountMgCl2 7 H2O 0.04 gCaCl2 H2O 0.13 gNaHCO3 0.42 g0.2 M KH

42、2PO4 7.70 mL0.2 M K2HPO4 12.3 mLNH4Cl 0.11 gKSCN 0.19 g(NH2)2CO 0.12 gNaCl 0.88 gKCl 1.04 gMucin 3.00 gDistilled water 1000 mLpH 7E2721 1637.1.1.1 The WHO Manual onAnimal Influenza Diagnosis and Surveillance contains specific procedures for preparing influenzavirus and titering samples. Appendix X2

43、also has specific information on titrating viable influenza viruses. Other viruses may beused, but conditions for propagation and enumeration are not provided in this method.7.1.2 MadinDarby Canine Kidney (MDCK) Cell Line, ATCC CRL-34.7.1.3 Artificial Saliva, see Table 1 in section 6.1.1.4.7.1.4 Min

44、imal Essential Medium With Earles Balanced Salts (EMEM).7.1.5 Heat-Inactivated Fetal Bovine Serum (45 min at 56C).7.1.6 Penicillin/Streptomycin, 10 000 units penicillin and 10 mg streptomycin per mL.7.1.7 L-Glutamine, 200 mM in 0.85 % NaCl.7.1.8 Crystal Violet.7.1.9 Glutaraldehyde.7.1.10 TPCKTrypsin

45、.7.1.11 Phosphate Buffered Saline (PBS).7.1.12 Bovine Serum Albumin.7.1.13 TrypsinEDTA Solution0.05 % trypsin, 0.53 mM EDTA in Hanks balanced salts solution without sodium bicarbonate,calcium, and magnesium.7.1.14 Distilled Water and Purified Water.7.1.15 Ethanol, laboratory grade.7.1.16 Bleach.7.2

46、MaterialsThe list is specific for influenza use. Other reagents may be needed if a different virus is used.7.2.1 Tissue Culture Treated FlasksT-75, T-175, 12-well, and 96-well plates.7.2.2 Pipettes, 1, 5, 10, and 25 mL.7.2.3 Test Tube Rack.7.2.4 Micropipettes, capable of delivering 0.001 mL accurate

47、ly and consistently.7.2.5 1.7-mL Sterile Microcentrifuge Tubes.7.2.6 15-mL Sterile Centrifuge Tubes.7.2.7 50-mL Sterile Centrifuge Tubes.7.2.8 Test Materials.8. Sampling, Test Specimens, and Test Units8.1 Cut test specimens from finished products or from specimens that can be documented as represent

48、ative of finished products.The configuration of the particular aerosol device dictates the size and type of each specimen. Place specimens into the dropletloader in the proper orientation. In some cases the complete finished product may be used, which obviates the need for cutting“coupons.”9. Experi

49、mental Design9.1 A minimum of three independent test and control samples must be evaluated so that fundamental statistical analysis of thedata can be performed.10. Test Procedure10.1 Apparatus OperationAppendix X1 describes a droplet loading device and details the standard protocols for operation ofthe device. General information that is independent of the droplet devices is listed below.10.2 Perform Neutralizer Effectiveness TestThe objective of this test is to determine whether toxic effects from the chemicalor physical decontaminati