ASTM E2755-2015 Standard Test Method for Determining the Bacteria-Eliminating Effectiveness of Healthcare Personnel Hand Rub Formulations Using Hands of Adults《利用成人手部测定医疗保健人员手搓配方灭菌.pdf

1、Designation: E2755 15Standard Test Method forDetermining the Bacteria-Eliminating Effectiveness ofHealthcare Personnel Hand Rub Formulations Using Handsof Adults1This standard is issued under the fixed designation E2755; the number immediately following the designation indicates the year oforiginal

2、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.1. Scope1.1 This test method is designed to determine the activity ofhealthcare

3、personnel hand rubs, (also known as hand rubs,hygienic hand rubs, hand sanitizers, or hand antiseptics)against transient microbial skin flora on the hands after a singleapplication and after repeated applications.1.2 Performance of this procedure requires the knowledgeof regulations pertaining to th

4、e protection of human subjects(see 21 CFR Parts 50 and 56).1.3 This test method should be performed by persons withtraining in microbiology, in facilities designed and equippedfor work with potentially infectious agents at biosafety level2.21.4 UnitsThe values stated in SI units are to be regardedas

5、 standard. No other units of measurement are included in thisstandard.1.5 This standard does not purport to address all of thesafety concerns, if any, associated with its use. It is theresponsibility of the user of this standard to establish appro-priate safety and health practices and determine the

6、 applica-bility of regulatory limitations prior to use. For more specificprecautionary statements, see 8.2.2. Referenced Documents2.1 ASTM Standards:3E1054 Test Methods for Evaluation of Inactivators of Anti-microbial AgentsE1174 Test Method for Evaluation of the Effectiveness ofHealth Care Personne

7、l Handwash FormulationsE2276 Test Method for Determining the Bacteria-Eliminating Effectiveness of Hygienic Handwash andHandrub Agents Using the Fingerpads of AdultsE2756 Terminology Relating to Antimicrobial and AntiviralAgents2.2 Other Standards:AATCC Test Method 147 2004 Antibacterial Activity As

8、-sessment of Textile Materials: Parallel Streak Method421 CFR Parts 50 and 56 Protection of Human Subjects;Institutional Review Boards53. Terminology3.1 Definitions: For definitions of terms used in thisdocument, see Terminology E2756.3.2 Definitions of Terms Specific to This Standard:3.2.1 healthca

9、re personnel handrub, nan antimicrobialgel, foam, liquid, spray, or wipe, applied by rubbing to reducethe transient microbial skin flora on hands that are not visiblysoiled, and which does not require a post-treatment water rinse.Such agents may also be referred to as hand rubs, hygienichand rubs, o

10、r hand antiseptics.3.2.2 healthcare personnel handwash, na cleanser orwaterless agent intended to reduce transient microbial skinflora on the hands.3.2.3 test bacteria, nan applied inoculum of bacteria thathas characteristics which allow it to be readily identified. Testbacteria are used to simulate

11、 a topical transient microbialcontaminant. This may also be referred to as a test organism,marker organism, simulant, or contaminant.3.2.4 test material, na product or formulation whichincorporates an antimicrobial ingredient(s).1This test method is under the jurisdiction of ASTM Committee E35 onPes

12、ticides, Antimicrobials, and Alternative Control Agents and is the directresponsibility of Subcommittee E35.15 on Antimicrobial Agents.Current edition approved May 15, 2015. Published June 2015. originallyapproved in 2010. Last previous edition approved in 2010 as E275510. DOI:10.1520/E2755152CDC-NI

13、H, Biosafety in Microbiological and Biomedical Laboratories, 5th ed.,U.S. Department of Health and Human Services, U.S. Government Printing Office,Washington, DC, 2007.3For referenced ASTM standards, visit the ASTM website, www.astm.org, orcontact ASTM Customer Service at serviceastm.org. For Annual

14、 Book of ASTMStandards volume information, refer to the standards Document Summary page onthe ASTM website.4Available from American Association of Textile Chemists and Colorists(AATCC), P.O. Box 12215, Research Triangle Park, NC 27709, http:/www.aatcc.org.5Available from U.S. Government Printing Off

15、ice Superintendent of Documents,732 N. Capitol St., NW, Mail Stop: SDE, Washington, DC 20401, http:/www.access.gpo.gov.Copyright ASTM International, 100 Barr Harbor Drive, PO Box C700, West Conshohocken, PA 19428-2959. United States14. Summary of Test Method4.1 This test method uses adult subjects w

16、ho have provideda written informed consent and whose hands have beendetermined to be free from any apparent damage at the time ofparticipation in the study. Subjects are to refrain from use ofany antimicrobials for at least one week prior to the initiationof the test procedure (see Section 11).4.2 S

17、ubjects hands are artificially contaminated with 0.2mL of a high-titer suspension of the test bacteria which isdistributed over all surfaces of the hands and fingers to producea minimum baseline recovery level of 108cfu/hand. BecauseSerratia marcescens is relatively sensitive to drying, the hightite

18、r suspension is prepared by growing in broth with vigorousaeration, followed by a 10-fold concentration with centrifuga-tion. Staphylococcus aureus is more resistant to drying and istherefore not concentrated after growth with vigorous aerationin broth.4.3 Test material effectiveness is measured by

19、comparingthe number of test bacteria recovered from contaminated handsafter use of the test material to the number recovered fromcontaminated hands not exposed to the test material. Activityof the test material is measured following a single applicationand after multiple consecutive contamination/ap

20、plicationcycles in a single day. Evaluating effectiveness after multipleapplications simulates repeated use of hand rubs in clinicalsettings and determines whether progressive build-up of non-volatile ingredients from the test material inhibits the antimi-crobial action. An abbreviated test measurin

21、g activity of thetest material following a single application may be used tosimulate situations where high frequency use is not expected.5. Significance and Use5.1 Hand hygiene is considered one of the most importantmeasures for preventing the spread of infectious microorgan-isms. Hand rubs reduce t

22、he microbial load on the handswithout the use of soap and water, and are thus an importanttool in the practice of good hand hygiene. Alcohol-based handrubs are recommended in healthcare settings for use on handsthat are not visibly soiled. They are formulated to be appliedfull strength to dry hands,

23、 “rubbed in” until dry, and are notrinsed off.5.2 This test method is designed specifically to evaluatehand rubs for efficacy in eliminating bacteria fromexperimentally-contaminated hands. It is designed as an alter-native to Test Method E1174, which was intended primarily toevaluate antimicrobial h

24、andwashing agents that are latheredwith the aid of water and then rinsed off. When using TestMethod E1174 to evaluate hand rubs, inadequate drying of thehands after contamination dilutes the test material and cancompromise activity, to result in an underestimation of effec-tiveness. Additionally, be

25、cause hand rubs are not rinsed afterproduct use, activity can be further degraded by build-up ofsoil from the contaminating broth and inactivated challengebacteria on the hands.5.2.1 In this method, application to the hands of a smallvolume of high-titer test bacteria suspension minimizes soilload s

26、uch that the skin is completely dry prior to application ofthe test material. Further, by applying the bacterial suspensiononly prior to those test material application cycles followed bysampling, excessive buildup of killed bacteria on the hands isavoided, and the potential impact of non-volatile t

27、est productingredients on bacteria-eliminating effectiveness after ten con-secutive applications can be specifically assessed.5.3 Areference control is evaluated for each subject prior toevaluation of the test material. Data from the reference controlhelps to control for inter-subject variability, i

28、nter-experimentalvariability, and inter-laboratory variability; and enables im-proved statistical comparison of test materials evaluated in thesame experiment.5.4 This test method can be used to test any form of handrubs, including gels, rinses, sprays, foams, and wipes whenused according to label d

29、irections at typical “in-use” doses.5.5 Susceptibility to biocides can vary among differentspecies of bacteria and major differences have been notedbetween gram-negative and gram-positive organisms. This testmethod provides the option to use either a gram-negativebacterium (Serratia marcescens) or a

30、 gram-positive bacterium(Staphylococcus aureus) as the test organism. S. marcescens isused as a test organism in both Test Method E1174 and TestMethod E2276. S. aureus is a highly relevant pathogen inhealthcare, institutional, and community settings. Moreover,hands are an important vehicle in the tr

31、ansfer of S. aureusbetween people and the environment, and in the transferbetween individuals.5.6 This test method may be used as an alternative to TestMethod E2276, which limits the test bacteria to the fingerpadsand does not incorporate actual use conditions such as frictionduring hand rubbing.5.7

32、 The investigator should be aware of potential healthrisks associated with the use of these organisms and precau-tions similar to those referenced in Section 8 should be taken.6. Apparatus6.1 CentrifugeFor the sedimentation of S. marcescens forconcentration.6.2 Centrifuge TubesSterile, for sedimenta

33、tion of S. marc-escens for concentration.6.3 Colony CounterAny of several types may be used; forexample, Quebec colony counters and similar devices.Automated, computerized plater/counter systems may also beused.6.4 GlovesSterile, loose-fitting, unlined, powder-freegloves possessing no antimicrobial

34、properties. Perform a zoneof inhibition test, such asAATCC Test Method 147, to evaluatethe antibacterial activity.6.5 Handwashing SinkSufficient in size to permit hand-washing without the touching of hands to sink surface or othersubjects.6.5.1 Water Faucet(s)Located above the sink at a heightto per

35、mit hands to be held higher than the elbow during thewashing procedure.E2755 1526.5.2 Tap Water Temperature Regulator and TemperatureMonitorTo set and maintain the tap water temperature at40 6 2C.6.6 IncubatorCapable of maintaining temperatures of35 6 2C and 25 6 2C. The latter temperature ensures a

36、d-equate pigment production for S. marcescens on solid media.6.7 Miscellaneous LabwareContinuously adjustable pi-petters (1-mL and 0.2-mL capacity) and sterile pipette tips,sterile serological pipettes (5.0-mL capacity), sterile culturetubes, sterile disposable Petri dishes, sterile syringes, Erlen-

37、meyer flasks, and beakers.6.8 Plastic BagsMay be used in place of gloves (6.4).Bags should be approximately 29 by 31 cm, possess noantimicrobial properties, and have a low bioburden. Perform azone of inhibition test, such as AATCC Test Method 147, toevaluate the antibacterial activity6.9 Sampling Co

38、ntainersSterile or sterilizable containershaving tight closures and sufficient capacity to hold 75 mLsampling solution (see 7.7).6.10 Shaking IncubatorRotary platform shaking incubatorcapable of maintaining 35 6 2C and capable of shaking at 250r/min. Alternatively, use an incubator capable of mainta

39、ining35 6 2C and able to accommodate a portable rotary shaker,capable of shaking at 250 r/min.6.11 SterilizerAny steam sterilizer capable of processingculture media and reagents.6.12 Timer (Stop-Clock)Type that can be read for minutesand seconds.6.13 TourniquetsChildrens size or any style capable of

40、securing gloves to the wrist.6.14 Vortex MixerAny vortex that will ensure propermixing of culture tubes.7. Reagents and Materials7.1 Antibiotic OintmentA topical, triple-antibiotic oint-ment for application to the hands after the final decontamina-tion.7.2 Cleansing WashA mild, proven non-antimicrob

41、ial liq-uid soap. May be purchased commercially or prepared accord-ing to the instructions provided in Test Method E1174.7.3 Chlorhexidine Skin CleanserAntiseptic skin cleansercontaining 4 % chlorhexidine gluconate (w/v) for hand decon-tamination.7.4 Culture Media:7.4.1 BrothSoybean-casein digest br

42、oth (tryptic soybroth) is recommended.7.4.2 Agar Plating Media:7.4.2.1 S. aureus Plating MediumHardyCHROM(trademark), Staph aureus, available from Hardy Diagnostics,is recommended. Other indicator media for S. aureus or MRSAmay be appropriate but should be validated prior to use.NOTE 1S. aureus form

43、s smooth, deep pink to fuchsia-colored colo-nies. The growth of most other organisms, including Staphylococcusepidermidis are partially to completely inhibited.7.4.2.2 S. marcescens Plating MediumSoybean-casein di-gest agar (tryptic soy agar) is recommended.7.5 Dilution FluidSterile Butterfields buf

44、fered phosphatediluent6(or other suitable diluent) adjusted to pH 7.2 6 0.1 andcontaining an effective inactivator for the test material, ifnecessary.NOTE 2Inactivator is only required if neutralization of the testmaterial cannot be achieved upon dilution into the sampling solution (see7.7).7.6 Etha

45、nol Solution70 % ethanol in water (v/v) for handdecontamination.7.7 Sampling SolutionDissolve 0.4 g KH2PO4, 10.1 gNa2HPO4, 1.0 g isooctylphenoxypolyethoxyethanol (forexample, Triton X-100), and appropriately validatedneutralizers, if necessary, in distilled water.Adjust pH to 7.8 60.1 with 0.1 N HCl

46、 or 0.1 N NaOH and bring volume to 1 Lwith distilled water. Sterilize in an autoclave and asepticallydispense 75-mL portions into sterile sampling containers (see6.9).7NOTE 3Aneutralizer validation should be conducted according toTestMethods prior to the study. Test Methods E1054 provides a list ofn

47、eutralizers appropriate for commonly used antimicrobial agents. In somecases (for example, some alcohol-based hand rubs) neutralization isachieved by dilution alone.7.8 Test MaterialUse directions provided with the testmaterial. If directions are not provided, use the directions givenin this method.

48、7.9 Reference Control60% isopropanol in water (v/v).8. Test Bacteria8.1 Serratia marcescens (ATCC 14756). This strain forms astable red pigmentation at 25C.8.2 Staphylococcus aureus (ATCC 6538 (methicillin-sensitive) or ATCC 33591 (methicillin-resistant) is an alterna-tive test bacteria. S. aureus i

49、s differentiated from residentmicrobial skin flora (including Staphylococcus epidermidis)with chromogenic indicator medium (see 7.4.2.1).(WarningApplication of microorganisms to the skin mayinvolve a health risk. Determine the antibiotic sensitivityprofile of the test bacteria prior to applying to the skin. Afterthe test has been completed, decontaminate the subjects handsand follow proper procedures to reduce infection risk (12.1 12.4). If an infection occurs, provide the antibiotic susceptibil-ity profile to the attending clinician