双重pcr诊断棘阿米巴角膜炎.研究

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1、温州医学院硕士学位论文StudyofutilitymultiplexPCRindiagnosisofAcanthamoebakeratitisI⋯Graduate：QINQianAdvisor：Prof．PANChang—wangProf．LIANGShao．huiProf．戳0Zhi．huaCollegeofClinicalLaboratoryMedicineo埘ective：Methods：AbstractAcanthamoebakeratitis(AK)isarefractor,progressive

2、，sightthreateningcomealdisease．Fortheabsenceofspecificityclinicalmanifestationintheearlydisease，thediagnosticmethodlackssensitiveandspecificity．SOthemisdiagnosisandmisseddiagnosisofAKisoften．ThisstudyappliedmultiplexPCRmethod，amplificated18srDNAand26srDNAa

3、tthesametime，toexploreanearlydiagnosisAKmethod，whichwasshortcut，，convenience，specificityandsensitivity．1．MorphologyandculturestudyofAcanthamoeba：Culturedacanthamoeba(GenetypeCJY／s，CJY／s2，Neff．YanbianMedicalCollegeoffered)withPYGfluid，keptspecies，observedth

4、emorphologychangeofAcanthamoeba’StrophczoiteandcystbyGramstaining．2．ToestablishamethodologyofPCRandMultiplexPCRamplificated18srDNAor(and)26srDNA。3。Toestablishananimal(rabbit)modelofAcanthamoebakeratitisandmakeuseofmu、ltiplexPCRindiagnosie：Injectedsuspensio

5、nofAcanthamoebagenetypeCYL／S2intotherabbit’leftcorneapropria，therighteyeascontrol，toestablishananimalmodelofAK．Observedthechangeoftherabbitleftandrighteyes．AppliedmultiplexPCRmethodtodetectAcanthamoebabydischargeofeye，rinsesolution，corneascraping，biopsy温确医

6、学院硕士学位论文tissue．GotrabbitcorneatopathologicalsectionandHEstaining，observedthemorphousofAcanthamoebatrophozoitesandcysts．4。ApplyMultiplexPCRtechnologyandNon—nutrientagarcultivationinclinical：Detected83caseovemightagingcontactlenssolutionofundergraduatestuden

7、tswhowearcontactlensandwithoutkeratitisclinicalmanifestation，40caseunusedcontactlenssolution，42casedischargeofeyesofdoubtfulAKpatientsand58casedischargeofeyesofhealthadult．RESULTS：1．Undermicroscope．themorphologydifferentwaslessamong3genetypesAcanthamoeba．G

8、enesequencingshowedAcanthamoebasp．CJY／sandCJY／s2belongtogeneT4type，A。castellaniiNeffbelongtoNefftype。Therewere99％genesamebetweenAcanthamoebasp。CJY／sandNeff．AfterGramstaining，brightdoublecircularityshapeofthec