ASTM E1838-2017 Standard Test Method for Determining the Virus-Eliminating Effectiveness of Hygienic Handwash and Handrub Agents Using the Fingerpads of Adults《测定采用成人指垫测定卫生洗手和擦手剂除病.pdf

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1、Designation: E1838 10E1838 17Standard Test Method forDetermining the Virus-Eliminating Effectiveness of HygienicHandwash and Handrub Agents Using the Fingerpads ofAdults1This standard is issued under the fixed designation E1838; the number immediately following the designation indicates the year ofo

2、riginal 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.INTRODUCTIONHands play an important role in the spread of many viruses.

3、Thus, proper and regular hand hygieneis crucial in preventing such spread, particularly in health-care settings, day-care centers, andfood-handling establishments. Many viruses that are known to spread through contaminated hands canremain infectious for several hours on human hands, and also may be

4、more resistant than the bacteriacommonly used to evaluate the microbicidal activity of handwash and handrub agents (1, 2, 3, 4).2Contaminated hands also can readily transfer infectious virus to other surfaces (1, 2, 3). Handantisepsis has been shown to interrupt the spread of viral infections (5, 6,

5、 7, 8, 9). Standardizedmethods This test method is to assess the virus-eliminating potential of handwash and handrub agentshave not in vivo.been available and this test method addresses the gap.1. Scope1.1 Human skin is not known to carry viruses as a part of its resident microbiota. microbiota, wit

6、h the notable exception ofpapilloma viruses (10). Hands transiently contaminated with viruses can, however, act as vehicles for the spread of many typesof viral infections. Hand hygiene is meant to reduce the load of viruses and other transient microorganisms on hands, therebyreducing the risk of di

7、sease transmission. Such reductions in the virus load may be due to a combination of virus inactivation andmechanical removal of infectious virus from the skin.1.2 This test method is designed to determine the comparative virus-eliminating effectiveness of microbicidal or non-microbicidal formulatio

8、ns. This test method is not meant for use with surgical hand scrubs or preoperative skin preps.NOTE 1The test method should be performed by persons with training in virology in facilities designed and equipped for work with infectious agentsat biosafety level 2 (611).1.3 The values stated in SI unit

9、s are to be regarded as standard. No other units of measurement are included in this standard.1.4 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 health pra

10、ctices and determine the applicability of regulatorylimitations prior to use.1.5 This international standard was developed in accordance with internationally recognized principles on standardizationestablished in the Decision on Principles for the Development of International Standards, Guides and R

11、ecommendations issuedby the World Trade Organization Technical Barriers to Trade (TBT) Committee.2. Referenced Documents2.1 ASTM Standards:3E2011 Test Method for Evaluation of Hygienic Handwash and Handrub Formulations for Virus-Eliminating Activity Using theEntire Hand1 This test method is under th

12、e jurisdiction of ASTM Committee E35 on Pesticides, Antimicrobials, and Alternative Control Agents and is the direct responsibility ofSubcommittee E35.15 on Antimicrobial Agents.Current edition approved April 1, 2010April 1, 2017. Published May 2010.June 2017. Originally approved in 1996. Last previ

13、ous edition approved in 20022010 asE1838 02.E1838 10. DOI: 10.1520/E1838-10.10.1520/E1838-17.2 The boldface numbers in parentheses refer to the list of references at the end of this standard.3 For referencedASTM standards, visit theASTM website, www.astm.org, or contactASTM Customer Service at servi

14、ceastm.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 standard and is intended only to provide the user of an ASTM standard an indication of what changes have been made to the previous version. B

15、ecauseit may not be technically possible to adequately depict all changes accurately, ASTM recommends that users consult prior editions as appropriate. In all cases only the current versionof the standard as published by ASTM is to be considered the official document.Copyright ASTM International, 10

16、0 Barr Harbor Drive, PO Box C700, West Conshohocken, PA 19428-2959. United States1E2276 Test Method for Determining the Bacteria-Eliminating Effectiveness of Hygienic Handwash and Handrub Agents Usingthe Fingerpads of AdultsE2613 Test Method for Determining Fungus-Eliminating Effectiveness of Hygien

17、ic Handwash and Handrub Agents UsingFingerpads of Adults3. Terminology3.1 Definitions of Terms Specific to This Standard:3.1.1 Health-care personnel (HCP), npersons who are directly related to provision of health care services. It includes all paidand unpaid persons working in health-care settings,

18、such as physicians, nurses, nursing assistants, therapists, technicians,emergency medical service personnel, dental personnel, pharmacists, laboratory personnel, autopsy personnel, students, trainees,and contractual staff, etc., who have the potential to get themselves exposed to patients and infect

19、ious materials.3.1.2 hygienic (health-care personnel) handwash agents, nagents generally used for handwashing by personnel in hospitals,other health-care facilities, day-care centers, nursing homes, and food-handling establishments should be safe for repeated use,nonirritating, fast-acting, and effi

20、cient in eliminating transient microorganisms from intact skin.3.1.3 nonmedicated soap, na soap or detergent that is mild to the skin and does not contain any germicidalmicrobicidalchemicals.3.1.4 soil(organic) load, na solution of one or more organic and/or inorganic substances added to the suspens

21、ion of the testorganism to simulate the presence of body secretions, excretions or other extraneous substances.3.1.5 virus-eliminating (killing/removing) agent, nany agent that rids hands of viruses by either killing them on the skin orby dislodging them for subsequent wash-off.3.1.6 virus inactivat

22、ing agent, nany agent that renders a virus noninfectious.4. Summary of Test Method4.1 This test method is conducted on a group of adult subjects who have provided informed consent and the skin of whose handshas been determined to be free from any apparent damage. The subjects are to refrain from usi

23、ng any products containingantimicrobial agents for at least one week prior to the test.Aknown volume of the test virus suspension is placed on a demarcatedarea on each fingerpad and the inoculum allowed to dry. The contaminated area then is exposed to the control (standard hard water)or test agent t

24、est or control agent or a vehicle (for example, standard hard water), and rubbed with a randomly chosen fingerpadfrom the opposite hand for the desired contact time and virus time. Virus remaining on the fingerpadfingerpads is then eluted andthe eluates titrated for infectious virus along with the r

25、equired controls. Percent and/orThe infectious units from the two thumbpadsor the pair of the fingerpads that were involved in a single treatment will be averaged. Percent or log10 reductions reductions, orboth, in the levels of infectious virus after treatment with the control and testtest or contr

26、ol agents are then determined. Thefingerpad method gives results that are comparable to those obtained using a whole-hand procedure (2, 7), anotherASTM standard(Test Method E2011). If two different formulations are being compared in the same test, one of them may be designated as areference and used

27、 in place of the hard water control. reference. If desired, one also may use tap water in parallel with the hardwater control to determine the influence of water hardness on the test products virus-eliminating activity.5. Significance and Use5.1 This in vivo procedure is designed to test the ability

28、 of hygienic handwash and handrub agents to reduce levels of selectedinfectious viruses from experimentally contaminated fingerpads of adults. Since the two thumbpads and all eight fingerpads canbe contaminated with virus and used in anya given test, it allows for the incorporation of input virus co

29、ntrol (two), virus remainingviable after the inoculum has been allowed to dry (two), virus eliminated after treatment with a control or reference solution (two),a wet inoculum input control, dried virus recovery control, and up to fourthree replicates to assess the virus-eliminating efficiencyof the

30、 substance under test. a test or control agent, or a vehicle material. No more than 100 L of the virus suspension are requiredto complete one test. The results of testing with this test method may form the basis for further tests using a suitable whole-handtest protocol (for example, Test Method E20

31、11).5.2 This test method is designed to be performed by a trained individual, who is responsible for choosing the appropriate hostsystem for the test virus and applying the techniques necessary for propagation and maintenance of host and test virus. For areference text, refer to Lennette et al (812)

32、.5.3 Whereas the method described here relates to testing with viruses of human origin, it can be readily adapted to work withanimal pathogenic viruses as well as bacteriophages. Standard methods for working with bacteria (Test Method E2276) and fungi(Test Method E2613) are also available.5.4 Infect

33、ious microorganisms left on hands after washing can be reduced further by drying the washed hands with paper, cloth,or warm air (913).Astep for the drying of fingerpads after exposure to the control or test substance,product, therefore, has not beenincluded to avoid virus removal by the drying proce

34、ss itself.5.5 This test method is not meant for use with surgical hand scrubs or preoperative skin preps.E1838 1725.6 The level of viable virus on each fingerpad after the drying of the inoculum in the dried inocula the control fingerpads shouldnot be less than 104 infectious units which would permi

35、t the detection of up to a 4 log10 reduction in the infectivity titer of thevirus by the test substanceproduct under the conditions of this test method.6. Equipment and Apparatus6.1 Laminar Flow CabinetA Class II biological safety cabinet is required for virus work. The procedures for the propermain

36、tenance and use of such cabinets are given in Ref (611).6.2 IncubatorAn incubator at 36 6 1C is needed for growing host cells and for incubating virus-infected cultures. If an opensystem is used for cell culture, a CO2 incubator will be required.6.3 Positive Displacement PipetteA pipette and pipette

37、 tips that accurately can dispense 10-L volumes.6.4 SterilizerAny steam sterilizer suitable for processing cell culture media and reagents is acceptable. The steam supplied tothe sterilizer must be free from additives toxic to cell cultures.6.5 Filter Sterilization SystemA membrane or cartridge filt

38、ration system (0.22-m pore diameter) is required for sterilizingheat-sensitive media and solutions.6.6 FreezersAfreezer at 2062C is required for the storage of fetal bovine serum and other additives for cell culture media.A second freezer at 70C or lower is required to store viruses6.7 RefrigeratorA

39、 refrigerator at 46 2C for storage of prepared cell culture media and reagents.6.8 TimerAny stopwatch that can be read in minutes and seconds.6.9 Magnetic Stirrer and MagnetsLarge enough to hold a 5-L beaker or Erlenmeyer flask for preparing cell culture media orother solutions.6.9 Handwashing SinkA

40、 sink of sufficient size to permit subjects to wash hands without touching hands to sink surface.6.9.1 Water Faucet(s), to be located above the sink at a height that permits the hands to be held higher than the elbow duringthe washing procedure. Faucets with electronic sensors or those that are wris

41、t-, elbow-, knee-, or foot-operated are preferred toavoid recontamination of the washed hands.6.9.2 Tap Water Temperature Regulator and Temperature Monitor, to monitor and regulate water temperature at 40 6 2C.6.10 Liquid Nitrogen Storage for CellsA proper liquid nitrogen container and liquid nitrog

42、en for cryopreservation of thestocks of cell lines.6.11 Inverted MicroscopeAn inverted microscope with 10 eye pieces and 5, 10, and 40 objectives.7. Materials and Reagents7.1 Serological PipettesSterile reusable or single-use pipettes of 10.0, 5.0, and 1.0-mL capacity.7.2 Cell Culture FlasksPlastic

43、flasks of 25 or 75-cm2 capacity for culturing cells and for preparing virus pools.NOTE 2Each flask for growing cell monolayers can be reused ten or more times before being discarded.NOTE 3Plastic cell culture ware may be purchased from most laboratory supply houses.7.3 Cell Culture Plates, 6-well2.0

44、 mL per well eluent (see 7.8) shall be added. This is used for virus elution from eachthumbpad and fingerpad.NOTE 4Alternatively, small dishes (for example, 35mm diameter) may be used for virus elution.7.4 Cell Culture Media and SupplementsCulture media and the types and ratios of supplements will v

45、ary depending on thecell line. Eagles minimal essential medium (EMEM) with 5 to 10 % fetal bovine serum (virus- and mycoplasma-tested) is usedfor growing a wide variety of cells (see Note 5).NOTE 5Materials and reagents for cell culture may be purchased from biological supply houses.7.5 Soil Load:7.

46、5.1 Bovine Serum, at a final concentration of 5 % in the virus inoculum (see Note 56).7.5.2 AYeast extract/BSA/Mucin tripartite soil load, as an alternative to serum.Add 0.5 g of tryptone or yeast extract to 10 mLof phosphate buffer. Add 0.5 g of bovine serum albumin (BSA) to 10 mL of phosphate buff

47、er. Add 0.04 g of bovine mucin to 10mLof phosphate buffer. Prepare the stock solutions separately and sterilize by passage through a 0.22 m pore diameter membranefilter, aliquot and store at either 462C or 2062C. To obtain a 500-L inoculum of the test inoculum, add to 340 L of themicrobial suspensio

48、n 25 L BSA, 100 L mucin and 35 L of tryptone yeast extract stock solutions. This mixture containsapproximately 2 g of total protein/L, which is approximately equivalent to the protein content of a 5 % solution of fetal bovineserum.NOTE 6Bovine serum is unsuitable for use as an organic load when work

49、ing with rotaviruses because of its rotavirus inhibitory andtrypsin-neutralizing activity.E1838 1737.6 Standard Hard WaterThe quality and disinfectant (for example, chlorine) residual in tap water can vary from site to siteand also at different times at the same site. The use of standard hard water, therefore, is recommended here to avoid variations inresults due to differences in tap water quality. Water Standard hard water prepared in accordance with AOAC 960.09 E andF (1014) toat a standard hardness of 200 ppm as calci