ASTM_E_1874_-_14.pdf

Designation:E187414Standard Test Method forRecovery of Microorganisms From Skin using the CupScrub Technique1This standard is issued under the fixed designation E1874;the number immediately following the designation 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.1.Scope1.1 This test method is designed to recover microorganismsfrom the skin of human subjects or human subject surrogates(animal skin,isolated porcine skin,human skin equivalents,and other such surfaces).1.2 Knowledge of microbiological techniques is requiredfor these procedures.1.3 It is the responsibility of the investigator to determine ifGood Laboratory Practice(GLP)and Good Clinical Practice(GCP)is required.1.4 The values stated in SI units are to be regarded asstandard.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 applica-bility of regulatory limitations prior to use.2.Referenced Documents2.1 ASTM Standards:2E1054 Test Methods for Evaluation of Inactivators of Anti-microbial Agents3.Terminology3.1 Definitions of Terms Specific to This Standard:3.1.1 contralateral,adjon or relating to the opposite side(of the body).3.1.2 resident flora,nmicroorganisms that live and mul-tiply on skin,forming a permanent population.3.1.3 scrub cups,nsterile cylinders of suitable composi-tion(that is,glass,ceramic,stainless steel,plastic,etc.)used toisolate a sample area of skin(or skin equivalent)and confine aaliquot of liquid which is used to facilitate the scrubbing of theskin and removal of microorganisms from the skin surface bypipetting.3.1.4 transient organisms,norganisms from the environ-ment that contaminate but do not normally colonize skin.4.Summary of Test Method4.1 This test method describes a technique suitable for therecovery of resident and transient microorganisms from humanor animal skin;the technique may be used in situ withinclinical protocols or in vitro for studies using isolated skin orskin equivalents.4.1.1 Resident microorganisms or transient microorganisms(previously applied to a test site),are recovered from the siteby pressing a rigid cylinder against the skin with sufficientpressure to form a seal and instilling recovery liquid into thecylinder.The surface of the skin is then mechanicallyscrubbed with a glass rod,rubber policeman,or some othersuitable device for a prescribed period of time.The fluid ispipetted from the cylinder into a test tube,or other suitablereceptacle,for further analysis.5.Significance and Use5.1 The procedure can be incorporated into protocols usedto evaluate test materials containing antibacterial ingredientsthat are intended to reduce significantly the number of organ-isms on intact skin.It also may be used to provide an indicationof residual antibacterial activity.Examples of test materials,forwhich this method is applicable,include hand-washes,surgicalscrubs,acne reduction products,and others.For each type oftest material,types of resident flora or transient organisms,ora combination thereof,may differ and should be considered(this is,aerobic bacteria,anaerobic bacteria,yeast,or mold).5.2 The procedure may be used in protocols intended toevaluate and identify resident flora from the skin.5.3 Performance of this technique may require the knowl-edge of regulations pertaining to the protection of humansubjects if the protocol involves application of the technique tothe skin of human subjects.1This tests method is under the jurisdiction of ASTM Committee E35 onPesticides,Antimicrobials,and Alternative Control Agents and is the directresponsibility of Subcommittee E35.15 on Antimicrobial Agents.Current edition approved April 1,2014.Published April 2014.Originallyapproved in 1997.Last previous edition approved in 2009 as E187509.DOI:10.1520/E1874-14.2For referenced ASTM standards,visit the ASTM website,www.astm.org,orcontact ASTM Customer Service at serviceastm.org.For Annual Book of ASTMStandards volume information,refer to the standards Document Summary page onthe ASTM website.Copyright ASTM International,100 Barr Harbor Drive,PO Box C700,West Conshohocken,PA 19428-2959.United States1 6.Apparatus6.1 SterilizerAny suitable steam sterilizer capable of pro-ducing the conditions of sterilization.7.Reagents and Materials7.1 Scrub CupsSterile cylinders of suitable composition,preferably with rod handles to facilitate stabilization,heightapproximately 2.5 cm,inside diameter of convenient size.Useful sizes range from approximately 1.5 to 4.0 cm.7.2 Polished Glass Rod or Rubber PolicemanCan befashioned in the laboratory or purchased.7.3 PipettorWith disposable tips to deliver appropriatevo