ASTM E1052-96(2002)

NOTICE: This standard has either been superseded and replaced by a new version or withdrawn.
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Designation:E1052–96 (Reapproved 2002)
Standard Test Method for
Efficacy of Antimicrobial Agents Against Viruses in
Suspension
This standard is issued under the fixed designation E1052; the number immediately following the designation indicates the year of
original adoption or, in the case of revision, the year of last revision.Anumber in parentheses indicates the year of last reapproval.A
superscript epsilon (´) indicates an editorial change since the last revision or reapproval.
1. Scope 2.2 Federal Standards:
Title 40, Code of Federal Regulations (CFR), Environmen-
1.1 This laboratory test method is a suspension test used to
talProtectionAgency,Part160,GoodLaboratoryPractice
evaluate the effectiveness of antimicrobial solutions against
Standard
specific viruses. This test method may be employed with most
Title 21, Code of Federal Regulations (CFR), Food and
viruses and is designed for cell culture host systems.
Drug Administration, Part 58, Laboratory Practice for
1.2 This test method should be performed only by those
Nonclinical Laboratory Studies
trained in microbiological or virological techniques.
1.3 This standard does not purport to address all of the
3. Summary of Test Method
safety concerns, if any, associated with its use. It is the
3.1 Onepartofthevirussuspensionisaddedtoninepartsof
responsibility of the user of this standard to establish appro-
theappropriatelydilutedantimicrobial.Thevirusisexposedto
priate safety and health practices and determine the applica-
the virucide for the length of time that is representative of
bility of regulatory limitations prior to use. The user should
actual use conditions or the label directions of the product (for
consult a reference for the laboratory safety recommenda-
example, from 15 sec for a handsoap to 10 min or longer for a
tions.
antimicrobial solution). The tests also should be performed at
1.4 It is the responsibility of the investigator to determine
the temperature most representative of actual use conditions
whether Good Laboratory Practice regulations (GLPs) are
(usually22 62°C).Thevirus-antimicrobialmixtureisassayed
required and to follow them where appropriate (40 CFR, Part
inahostsystemappropriateforthetestvirus.Thevirustiterof
160 for EPA submissions andCFR, Part 58 for FDA submis-
the stock virus is determined by the median cell culture
sions). Refer to the appropriate regulatory agency for perfor-
infective dose (CCID ), plaque assay or other quantifiable
mance standards of virucidal efficacy.
measure of infectivity. Cytotoxicity to the host system (from
2. Referenced Documents the antimicrobial) at the tested concentration also is deter-
mined.Thevirus-antimicrobialmixtureisassayedinnumerous
2.1 ASTM Standards:
units of the host system at a dilution just beyond the cytotoxic
E1053 Test Method for Efficacy of Virucidal Agents In-
range of the antimicrobial. At least three replicate determina-
tended for Inanimate Environmental Surfaces
tions are performed on controls and experimentals to confirm
E1153 Test Method for Efficacy of Sanitizers Recom-
virus inactivation by a batch of antimicrobial. Results are
mended for Inanimate Non-Food Contact Surfaces
recorded as the median value of log -virus inactivation.
E1482 Test Method for Neutralization of Virucidal Agents 10
3.2 This test method is designed to be performed by a
in Virucidal Efficacy Evaluations
trained microbiologist or virologist who is responsible for
choosing the appropriate host system for the test virus, and
This test method is under the jurisdiction of ASTM Committee E35 on
applying the techniques necessary for propagation and main-
PesticidesandisthedirectresponsibilityofSubcommitteeE35.15onAntimicrobial
tenance of host and test virus. For a reference text, refer to
Agents.
Schmidt and Emmons.
Current edition approved Oct. 10, 1996. Published December 1996. Originally
published as E1052–85. Last previous edition E1052–85 (1990). DOI: 10.1520/
E1052-96R02.
CDC-NIH, Biosafety in Microbiological and Biomedical Laboratories, Third
Edition, U.S. Department of Health and Human Services, Washington, DC, May
1993. Available from U.S. Government Printing Office, Superintendent of Docu-
For referenced ASTM standards, visit the ASTM website, www.astm.org, or ments, Washington, DC 20402.
contact ASTM Customer Service at service@astm.org. For Annual Book of ASTM Diagnostic Procedures for Viral, Rickettsial and Chlamydial Infections,N.J.
Standards volume information, refer to the standard’s Document Summary page on Schmidt and W. W. Emmons, Eds, Sixth Edition, Amer. Pub. Hlth. Assoc.,
the ASTM website. Washington, DC 1989.
Copyright © ASTM International, 100 Barr Harbor Drive, PO Box C700, West Conshohocken, PA 19428-2959, United States.
E1052–96 (2002)
4. Significance and Use (representative of those most easily inactivated) and an aden-
ovirus (representative of intermediate resistance to germi-
4.1 This test method is to be used to determine the effec-
cides).Thefollowingisalistofsuggestedvirusstrainsthatare
tiveness of antimicrobial solutions against designated proto-
typically assayed, as well as cell cultures that support their
type viruses that are in suspension.
growth.
4.2 The effective antimicrobial concentration should be
6.3 Suggested test virus strains and cell cultures.
determined using cell cultures as the host system for specific
6.3.1 Poliovirus, Type 1, Chat strain, American Type Cul-
viruses.
ture Collection (ATCC) VR 192. Cell line options: Monkey
4.3 This suspension test is for special applications of viru-
KidneyCells(VERO);HumanEpidermoidCarcinoma,Larynx
cides, such as inactivation of viruses in contaminated liquid
(HEp-2); African Green Monkey Kidney (CV-1).
wastes,andasafirststageindeterminingvirucidalpotentialof
6.3.2 Hepatitis A Virus, HM-175 strain, ATCC VR-2093.
liquidchemicalgermicides,liquidhandsoaps,OTCtopicalsor
Cell line options: Fetal Kidney, Rhesus Monkey, Continuous
other skin products. Regulatory agencies may require addi-
(FRhK-4).
tional tests to demonstrate overall virucidal activity.
6.3.3 Herpes simplex, Type 1, strain F (1), ATCC VR-733.
Cell line options: VERO, HEp-2.
5. Materials and Reagents
6.3.4 Cytomegalovirus, strainAD-169,ATCCVR-538. Cell
5.1 Cell Culture Technique.
line options: Human Diploid Lung (MRC-5 or WI-38).
5.1.1 Cell Culture System appropriate for test virus.
6.3.5 Adenovirus, Type 2, Adenoid 6 strain, ATCC VR-2.
5.1.2 Growth Media/Maintenance Media, Medium 199, Ea-
Cell line options: Human Lung Carcinoma (A549), Hep-2.
gle’s minimal essential medium (EMEM) or equivalent,
6.3.6 Influenza A , Hong Kong Strain,ATCC VR-544. Cell
supplemented with appropriate concentration of serum (inac-
line options: Canine Kidney (MDCK); Rhesus Monkey Cells,
tivated and mycoplasma-free), antibiotics and other growth
Continuous (LLC-MK2).
factors as needed.
6.3.7 Respiratory Syncytial Virus, Long strain, ATCC VR-
5.1.3 Diluent,The media listed in 5.1.2, phosphate buffered
26. Cell line options: HEp-2, MRC-5.
saline, trypticase soy b
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