猪多杀性巴氏杆菌(PM)PCR诊断技术的建立-豆柴文库

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猪多杀性巴氏杆菌(PM)PCR诊断技术的建立
Title:EstablishmentofPorcineMulticentricLymphoma(PM)PCRDiagnosticTechnology
Abstract:
PorcineMulticentricLymphoma(PM)isafatalneoplasticdiseaseaffectingpigsworldwide.Currently,thereisalackofeffectivediagnostictoolstodetectPMatanearlystage.Inthisstudy,weaimedtoestablishaPolymeraseChainReaction(PCR)diagnostictechnologyfortheaccuratedetectionofPM.WedevelopedspecificprimerstargetingtheBacillusanthracisbacteria,thecausativeagentofPM,andoptimizedthePCRconditionsforhighsensitivityandspecificity.OurresultsdemonstratethepotentialofthisPCR-baseddiagnostictechniqueinimprovingtheearlydetectionandcontrolofPMinpigs.
Introduction:
PorcineMulticentricLymphoma(PM)isamalignantlymphoproliferativedisorderthatprimarilyaffectspigs.PMischaracterizedbytheuncontrolledgrowthoflymphocytes,leadingtotheformationoftumormassesinvariousorgans,includinglymphnodes,spleen,liver,andkidneys.Thediseaseoftengoesundetecteduntiladvancedstages,resultinginsignificanteconomiclossesforpigfarmers.EarlydiagnosisofPMiscriticalforeffectivecontrolandpreventionstrategies.However,currentdiagnosticmethods,suchashistopathology,lacksensitivity,specificity,andturnaroundtime.Therefore,thedevelopmentofarapidandaccuratediagnostictechnique,suchasPCR,isofutmostimportancetocombatthisdisease.
MaterialsandMethods:
1.BacterialIsolationandCulture:
-BacillusanthracisstrainsisolatedfromPM-infectedpigswereobtained.
-Isolateswereculturedinappropriatemediaandconditionstoobtainpurebacterialcultures.
2.PrimerDesign:
-SpecificprimersweredesignedtargetingconservedregionsoftheBacillusanthracisgenomeassociatedwithPM.
-Primersweredesignedusingbioinformaticstoolsandcheckedforspecificityusingsequencedatabases.
3.PCROptimization:
-VariousPCRconditionsweretested,includingprimerconcentration,annealingtemperature,PCRcyclenumber,andDNAtemplateconcentration.
-Optimalconditionsyieldingthehighestsensitivityandspecificitywereselected.
4.SensitivityandSpecificityTesting:
-TestsamplesincludedknownPM-positiveandPM-negativepigtissues.