实时荧光PCR法检测动物源性成分的定性方法验证-豆柴文库

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实时荧光PCR法检测动物源性成分的定性方法验证
Title:ValidationofaQualitativeReal-TimePCRMethodfortheDetectionofAnimal-DerivedIngredients
Abstract:
Inrecentyears,therehasbeenanincreaseddemandforfoodproductsthatarefreefromanimal-derivedingredients,duetovariousreasonssuchasreligiousdietaryrestrictions,ethicalconcerns,andhealthpreferences.Toensuretheaccuracyoffoodlabelingandtomeetconsumerexpectations,itisessentialtodeveloprobustandreliablemethodsforthequalitativedetectionofanimal-derivedingredients.Thispaperaimstovalidateaqualitativereal-timepolymerasechainreaction(PCR)methodforthedetectionofanimal-derivedingredientsinfoodproducts.
Introduction:
Theaccurateandreliabledetectionofanimal-derivedingredientsisessentialforconsumerswhofollowspecificdietaryrestrictionsorhavepersonalpreferences.Traditionalmethodsforthedetectionofanimal-derivedingredients,suchasmicroscopicexaminationandimmunoassays,aretime-consuming,labor-intensive,andmaylackspecificity.Real-timePCRhasemergedasapromisingtechniqueforthedetectionandquantificationoftargetDNAsequencesduetoitssensitivity,specificity,speed,andpotentialforautomation.
MaterialsandMethods:
1.Designofprimersandprobes:PrimersandprobesspecifictotargetanimalDNAsequencesweredesignedusingavailablegenomicdatabases.
2.DNAextraction:DNAextractionwasperformedusingacommercialDNAextractionkitfollowingthemanufacturer'sinstructions.
3.Real-timePCRamplification:Real-timePCRamplificationwascarriedoutusingareal-timePCRinstrumentwiththedesignedprimersandprobes.Thereactionconditionsandcyclingparameterswereoptimizedformaximumsensitivityandspecificity.
4.Amplificationcurveanalysis:Theamplificationcurvesobtainedfromreal-timePCRwereanalyzedtodeterminethepresenceorabsenceofthetargetanimalDNAinthesample.
5.Sensitivityandspecificityevaluation:Thesensitivityandspecificityofthereal-timePCRmethodwereevaluatedbytestingknownpositiveandnegativecontrolsamples,aswellasblindsamplesobtainedfromfoodproducts.
Results:
ThedesignedprimersandprobessuccessfullyamplifiedthetargetanimalDNAsequenceswithh