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小麦和榛子过敏原成分检测的实时荧光PCR方法.docx

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小麦和榛子过敏原成分检测的实时荧光PCR方法 Title:Real-timeFluorescentPCRMethodforDetectionofAllergenicComponentsinWheatandHazelnutAllergens Abstract: Allergicreactionstofoodareagrowingconcernworldwide,affectingasignificantportionofthepopulation.Wheatandhazelnutareamongthemostcommonfoodallergens,causingadversereactionsrangingfrommilddiscomforttosevereanaphylaxis.Accuratedetectionandquantificationoftheseallergeniccomponentsareessentialforensuringfoodsafetyandconsumerprotection.Thispaperpresentsacomprehensivereviewofthereal-timefluorescentpolymerasechainreaction(PCR)methoddevelopedfortheidentificationandquantificationofwheatandhazelnutallergensinfoodproducts. 1.Introduction: Foodallergieshavebecomeamajorpublichealthissue,necessitatingtheimplementationofeffectivedetectionmethodsforallergeniccomponentsinfood.Withtheadventofmolecularbiologytechniques,real-timefluorescentPCRhasemergedasapowerfultoolforthedetectionandquantificationofallergeniccomponents.Thispaperfocusesonthedevelopmentandapplicationofreal-timefluorescentPCRforthedetectionofwheatandhazelnutallergens. 2.Background: 2.1Wheatallergens:Wheatallergycanmanifestasvariousallergicconditions,suchaseczema,allergicrhinitis,oranaphylaxis.Themajorwheatallergensincludeglutenproteins(gliadinsandglutenins)andnon-glutenproteins(amylase/trypsininhibitors,lipidtransferproteins,andalpha-amylaseinhibitors).Real-timefluorescentPCRallowsforthespecificamplificationandquantificationoftheseallergeniccomponents. 2.2Hazelnutallergens:Hazelnutallergyisoneofthemostcommontreenutallergies,oftencausingsevereallergicreactions.Themajorhazelnutallergensincludecora1,cora9,andcora14.Real-timefluorescentPCRcanaccuratelyidentifyandmeasurethepresenceoftheseallergeniccomponentsinfoodsamples. 3.Real-timeFluorescentPCRMethod: 3.1DNAextraction:Thefirststepinthereal-timefluorescentPCRmethodistheextractionofDNAfromthefoodsample.Variousmethods,suchasCTAB-basedextractionorcommercialDNAextractionkits,canbeemployeddependingonthesampletypeanddesiredDNAyield. 3.2Primerdesign:Speci