ISO TR 4752 2025.docx

ISOTechnicalReportISOTR4752Firstedition2025-06BiotechnologyInventoryofmethodsfordetectionofmicrobiologicalcontaminationinmammaliancellcultureBiotechnologieInventairedesmethodespourIadetectiondelacontaminationmicrobiologiquedanslacultureCeIIulairedemammiferesReferencenumberISO/TR4752:2025(en)COPYRIGHTPROTECTEDDOCUMENT©ISO2025Allrightsreserved.Unlessotherwisespecified,orrequiredinthecontextofitsimplementation,nopartofthispublicationmaybereproducedorutilizedotherwiseinanyformorbyanymeans,electronicormechanical,includingphotocopying,orpostingontheinternetoranintranet,withoutpriorwrittenpermission.PermissioncanberequestedfromeitherISOattheaddressbeloworISO,smemberbodyinthecountryoftherequester.ISOcopyrightofficeCP401Ch.deBlandonnet8CH-1214Vernier,GenevaPhone:+41227490111Email:copyrightiso.orgWebsite:www.iso.orgPublishedinSwitzerlandContentsPageForewordivIntroductionv1 Scope12 Normativereferences13 Termsanddefinitions14 Generalconcepts34.1 Introduction34.2 Detectionprocess35 Contaminationtestinformation46 Criticalcontrolpointsindetection56.1 Samples56.2 ReagentsandEquipment56.3 Operation66.4 Personnel66.5 Environment6Annex A (informative)Availableandexemplarytestingmethodsforbacteriaandfungi7Annex B (informative)AvailableandexemplarytestingmethodsforViruses12Annex C (informative)Availableandexemplarytestingmethodsformycoplasma15Bibliography18ForewordISO(theInternationalOrganizationforStandardization)isaworldwidefederationofnationalstandardsbodies(ISOmemberbodies).TheworkofpreparingInternationalStandardsisnormallycarriedoutthroughISOtechnicalcommittees.Eachmemberbodyinterestedinasubjectforwhichatechnicalcommitteehasbeenestablishedhastherighttoberepresentedonthatcommittee.Internationalorganizations,governmentalandnon-governmental,inliaisonwithISO,alsotakepartinthework.ISOcollaboratescloselywiththeInternationalElectrotechnicalCommission(IEC)onallmattersofelectrotechnicalstandardization.TheproceduresusedtodevelopthisdocumentandthoseintendedforitsfurthermaintenancearedescribedintheISO/IECDirectives,Part1.Inparticular,thedifferentapprovalcriterianeededforthedifferenttypesofISOdocumentshouldbenoted.ThisdocumentwasdraftedinaccordancewiththeeditorialrulesoftheISO/IECDirectives,Part2(seeWWW.iso.org/directives).ISOdrawsattentiontothepossibilitythattheimplementationofthisdocumentmayinvolvetheuseof（八）patent(三).ISOtakesnopositionconcerningtheevidence,validityorapplicabilityofanyclaimedpatentrightsinrespectthereof.Asofthedateofpublicationofthisdocument,ISOhadnotreceivednoticeof（八）patent(三)whichmayberequiredtoimplementthisdocument.HoweverlImplementersarecautionedthatthismaynotrepresentthelatestinformation,whichmaybeobtainedfromthepatentdatabaseavailableatWWW.iso.org/patents.ISOshallnotbeheldresponsibleforidentifyinganyorallsuchpatentrights.Anytradenameusedinthisdocumentisinformationgivenfortheconvenienceofusersanddoesnotconstituteanendorsement.Foranexplanationofthevoluntarynatureofstandards,themeaningofISOspecifictermsandexpressionsrelatedtoconformityassessment,aswellasinformationaboutISO'sadherencetotheWorldTradeOrganization(WTO)principlesintheTechnicalBarrierstoTrade(TBT工seeWWW.iso.org/iso/foreword.htmLThedocumentwaspreparedbyTechnicalCommitteeISO/TC276,Biotechnology,SubcommitteeSCLAnalyticalmethods.Anyfeedbackorquestionsonthisdocumentshouldbedirectedtotheuser'snationalstandardsbody.Acompletelistingofthesebodiescanbefoundatwww.iso.org/members.html.IntroductionCellcultureplaysanextremelyimportantroleinbiotechnologyandlifescienceresearch.Therefore,thereliabilityofscientificdatafromcellcultureisessential.Oneofthemostcommonproblemsincellcultureismicrobiologicalcontamination.Thisusuallyresultsincatastrophiccelldeathorcontaminationsustainedatlowlevels(sometimesduetotheuseofantibiotics),andthusimpactsthereliabilityofexperimentaldataderivedfromcellculture.Nonetheless,despiteitsprevalenceandimportance,thelackofopendiscussiondiscouragesdevelopmentandlearningofbestpracticestoavoidmicrobialcontaminationincellculture.Microbialcontaminationisaneconomicissueasitaffectsthereproducibilityofscientificdata,theefficiencyofinvitrocellcultureworkandmostimportantly,thesafetyoflaboratoryoperators.Itisthereforenecessarytomonitormicrobialcontaminationthroughafullunderstandingofthesourceofcontaminationandemploygoodtestingtechniquesbeforeandduringcellculture.Whenculturedcellshavebeencontaminated,firstthevariousaspectsofthecontaminationareidentified,includingthenatureofcontamination,thetimeofcontaminationandtheoperatingenvironment.Contaminantscanincludebacteria,fungi,mycoplasma,virusesandothertypesoforganisms.Inordertodemonstratetheabsenceofmicrobiologicalcontaminationincellculture,itcanbenecessarytoconductaseriesoftestsforlikelyorganismsandsuchanapproachwillbenefitfromriskassessmentofthecelltype,celloriginandreagentsusedfortheirculture.Environmentalmonitoringinareaswherecellsareculturedandstoredcan