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1、EN18057June2025EuropeanstandardNormeeuropeenneEuropaischenormICS67.120.10EnglishVersionFoodauthenticity-QuantitationofroedeerDNArelativetomammalianDNAinmeatandmeatproductsbyreal-timePCRAuthenticitedesaliments-QuantificationdeADNdechevreuilparrapportaADNdemammiferedanslaviandeetlesproduitscarriesparP
2、CRentempsreelLebensmittelauthentizitat-QuantifizierungvonReh-DNAimVerhaltniszuSaugetier-DNAinFleischundFleischproduktenmittelsReal-time-PCRThisEuropeanStandardwasapprovedbyCENon4May2025.CENmembersareboundtocomplywiththeCEN/CENELECInternalRegulationswhichstipulatetheconditionsforgivingthisEuropeanSta
3、ndardthestatusofanationalstandardwithoutanyalteration.Up-to-datelistsandbibliographicalreferencesconcerningsuchnationalstandardsmaybeobtainedonapplicationtotheCEN-CENELECManagementCentreortoanyCENmember.ThisEuropeanStandardexistsinthreeofficialversions(English,French,German).Aversioninanyotherlangua
4、gemadebytranslationundertheresponsibilityofaCENmemberintoitsownlanguageandnotifiedtotheCEN-CENELECManagementCentrehasthesamestatusastheofficialversions.CENmembersarethenationalstandardsbodiesofAustria,Belgium,Bulgaria,Croatia,Cyprus,CzechRepublic,Denmark,Estonia,Finland,France,Germany,Greece,Hungary
5、,Iceland,Ireland,Italy,Latvia,Lithuania,Luxembourg,Malta,Netherlands,Norway,Poland,Portugal,RepublicofNorthMacedonia,Romania,Serbia,Slovakia,Slovenia,Spain,Sweden,Switzerland,TiirkiyeandUnitedKingdom.EUROPEANCOMMITTEEFORSTANDARDIZATIONCOMITEEUROPEENDENORMALISATIONEuropaischeskomiteefurnormungCEN-CEN
6、ELECManagementCentre:RuedelaScience23,B-1040Brussels2025CENAllrightsofexploitationinanyformandbyanymeansreservedRef.No.EN18057:2025EworldwideforCENnationalMembers.ContentsPageEuropeanforeword3Introduction41 Scope52 Normativereferences53 Termsanddefinitions54 Symbolsandabbreviations65 Principle66 Rea
7、gentsandmaterials67 Apparatus88 Procedure88.1 Preparationofthetestsampleandtestportion88.2 PreparationofDNAextracts88.3 Preparationofroedeerandmammaliancalibrationstandards88.4 PCRsetup98.4.1 Samplesandcontrols98.4.2 Reactionmixes98.4.3 Thermalcycling109 Accept/rejectcriteria109.1 General109.2 Dataa
8、nalysis1110 Validationstatusandperformancecriteria1110.1 General1110.2 Repeatability1410.3 Reproducibility1410.4 Recovery1410.5 LimitofDetection(LOD)1410.6 Specificity1511 Testreport15Bibliography16EuropeanforewordThisdocument(ENl8057:2025)hasbeenpreparedbyTechnicalCommitteeCEN/TC460Foodauthenticity
9、”,thesecretariatofwhichisheldbyDIN.ThisEuropeanStandardshallbegiventhestatusofanationalstandard,eitherbypublicationofanidenticaltextorbyendorsement,atthelatestbyDecember2025,andcon11ictingnationalstandardsshallbewithdrawnatthelatestbyDecember2025.Attentionisdrawntothepossibilitythatsomeoftheelements
10、ofthisdocumentmaybethesubjectofpatentrights.CENshallnotbeheldresponsibleforidentifyinganyorallsuchpatentrights.Anyfeedbackandquestionsonthisdocumentshouldbedirectedtotheusersnationalstandardsbody.AcompletelistingofthesebodiescanbefoundontheCENwebsite.AccordingtotheCEN-CENELECInternalRegulations,then
11、ationalstandardsorganisationsofthefollowingcountriesareboundtoimplementthisEuropeanStandard:Austria,Belgium,Bulgaria,Croatia,Cyprus,CzechRepublic,Denmark,Estonia,Finland,France,Germany,Greece,Hungary,Iceland,Ireland,Italy,Latvia,Lithuania,Luxembourg,Malta,Netherlands,Norway,Poland,Portugal,Republico
12、fNorthMacedonia,Romania,Serbia,Slovakia,Slovenia7Spain,Sweden,Switzerland,TiirkiyeandtheUnitedKingdom.IntroductionFoodauthenticityandintegrityarekeyaspectsintermsofconsumerprotection.Sincethelate1980s,globalizationhastakenplaceinthetradeoffood.Duringthelastdecades,manyofmethodsapplyingPCR,particular
13、lyreal-timePCRprotocols,havebeenestablishedfortheidentificationofanimalspeciesusedforfoodconsumption(e.g.pig,cattle,sheep,horse,chickenandturkey).Gamemeatisparticularlysusceptibletofraudulentlabellingsinceitismorevaluablethanmeatfromdomesticanimals.Avarietyofgamemeatproducts,commonlycontainingreddee
14、rorroedeer,iscommerciallyavailable.Forverifyingthecorrectlabellingofcommercialgamemeatproducts,thedevelopmentofharmonizedandstandardizedprotocolsfortheauthenticationofmeatproductsisimportanttoestablishreliablemethodsforthedetectionofpotentialfoodfraud.1 ScopeThisdocumentspecifiesareal-timePCRprocedu
15、reforthequantitationoftheamountofroedeerDNArelativetototalmammalianDNAinmeatandmeatproducts.Resultsofthisroedeerassayareexpressedintermsofroedeer(CaPreOlUSCaPreOlUS)haploidgenomecopynumbersrelativetototalmammalianhaploidgenomecopynumbers.Thecontentofroedeercanalsobeexpressedasmassfractionin%usinggra
16、vimetricallypreparedcalibrationmaterialfrommeatmixturesormodelsamples.ThemethodhasbeenpreviouslyvalidatedinacollaborativestudyandappliedtoDNAextractedfromsamplesthatconsistofrawroedeermeatinarawpigmeatbackgroundaswellasrawandboiledsausages.ThelimitofdetectionoftheroedeerPCRhasbeendeterminedexperimentallytobeatleast5t
