Designation:E1536–00StandardPracticeforDetectionofMycoplasmaContaminationofBovineSerumbytheLargeVolumeMethod1ThisstandardisissuedunderthefixeddesignationE1536;thenumberimmediatelyfollowingthedesignationindicatestheyearoforiginaladoptionor,inthecaseofrevision,theyearoflastrevision.Anumberinparenthesesindicatestheyearoflastreapproval.Asuperscriptepsilon(e)indicatesaneditorialchangesincethelastrevisionorreapproval.1.Scope1.1Thispracticecoverstheproceduresusedfordetectionofmycoplasmacontaminationinserumbydirectmicrobiologicalculture.1.2Thispracticedoesnotcoverproceduresusedfordetec-tionofmycoplasmaincellcultures.1.3Thispracticedoesnotcoverindirectmethodsfordetectionofmycoplasmacontamination.1.4Thispracticedoesnotcovermethodsforidentificationofmycoplasmacultures.1.5Thisstandarddoesnotpurporttoaddressallofthesafetyconcerns,ifany,associatedwithitsuse.Itistheresponsibilityoftheuserofthisstandardtoestablishappro-priatesafetyandhealthpracticesanddeterminetheapplica-bilityofregulatorylimitationspriortouse.2.ReferencedDocuments2.1ASTMStandards:E1531PracticefortheDetectionofMycoplasmaContami-nationofCellCulturesbyGrowthonAgaroseMedium2E1532PracticefortheDetectionofMycoplasmaContami-nationofCellCulturesbytheUseoftheBisbenzamideDNA-BindingFluorochrome2E1533PracticeforIndirectDetectionofMycoplasmainCellCultureby48-6-Diamidino-2-2Phenylindole(DAPI)Staining23.Terminology3.1Definitions:3.1.1directmycoplasmadetection,n—demonstrationofcharacteristiccolonialgrowthonaxenicagarmedium.3.1.2largevolumetesting,n—usingalargevolumeinocu-luminanenrichmentculture.3.1.3mycoplasma(Mollicute),n—smallestprokaryotesca-pableofselfreplication.4.SignificanceandUse4.1Mycoplasmasofbovineoriginareprevalentcontami-nantsofcellcultures.Contaminationcanbedetectedbythelargevolumemethod.3,44.2Heatinactivatedserumneednotbetestedformycoplas-mas.Heatingserumto56°Cfor30minuteswillkillmyco-plasmas.4.3Mycoplasmasmaybepresentinanyparticularlotofserumbutmaynotbedetectedbecauseofinadequatesamplesize;thus,negativet...
