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ID:62180045
大小:1.55 MB
页数:39页
时间:2021-04-20
《最新肺炎支原体巢式PCR的优化教学讲义ppt课件.ppt》由会员上传分享,免费在线阅读,更多相关内容在教育资源-天天文库。
1、肺炎支原体巢式PCR的优化Mycoplasmapneumoniaeisafrequentcauseofcommunity-acquiredpneumoniafor10-40%inchildrenandadults.BecauseofthetreatmentofM.pneumoniainfectionwithβ-lactamantibioticsisineffectiveandtheclinicalmanifestationsofM.pneumoniaeinfectionarecomplicatedandnonspecific,soarapid,sensitiveandspecifi
2、claboratorytestisvitalforearlydiagnosisofM.pneumoniaeinfection.ConventionaltestsfordetectingM.pneumoniaehavetheirlimitations.IntroductionIntroductionSeveralPCR-relatedmethodsprovideenhancedsensitivityandhavebeensuccessfullyappliedforresearchpurposessuchasnestedPCR.TheP1adhesiongeneandthe16SrRN
3、AgenehavebeenutilizedwidelyinPCRtechniquesasthetargetsfordetectionofM.pneumoniae.Inthisstudy,wesoughttoidentifythemoresensitiveandspecifictarget(P1or16SrRNA)inM.pneumoniaedetectionandtoevaluatetheuseofnestedPCRforthediagnosisofMPinfectionfrompatientsinwhomM.pneumoniaewassuspected.NestedPCRsens
4、itivitytestsensitivitiesofnestedPCRforM.pneumoniae:LaneM:DNAmarker.Lane1:20ngofM.pneumoniaeFHstrain;Lane2:10ng;Lane3:10-1ng;Lane4:10-2ng;Lane5:10-3ng;Lane6:10-4ng;Lane7:10-5ng;Lane8:10-6ng;Lane9:negativecontrol.M123456789P1gene:1pg12345678912345678916SrRNA;0.1pgDetectionofclinicalsamplesP1adhe
5、siongene:(25/55;43.6%)Detectionofclinicalsamples16SrRNAgene(30/55;56.3%)discussionWiththedevelopmentofmolecularbiologytechniques,PCRtechnologyhasbecomethemostvaluablemethodforrapiddiagnosisofMycoplasmapneumoniaeinfection.BothP1adhesiongeneand16SrRNAgenearewidelyusedastargetsfordetectionofM.pne
6、umoniaebyPCR.However,itisstillinconclusiveforwhichtargetisbetterandoureffortistofindthemostsuitableone.Inthisstudy,wejointedorthogonalexperimentandsinglefactorteststooptimizeseveralcrucialconditionsofnestedPCRandfinallyconcludedtheoptimumreactionconditionsofthetwotargets.Thenwedetectedthesensi
7、tivityofthetwotargetsonthebasisoftheoptimalconditionsandtheresultsshowedthatthe16SrRNAgeneismoresensitivethantheP1adhesiongene.Wealsopresentedastudybyusingclinicalspecimensfromadultpatientsandfoundthat16SrRNAgenehasahigherpositiverateth
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