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1、Polymerasechainreaction–baseddetectionofPrhinovirus,respiratorysyncytialvirus,andcoronavirusinotitismediawitheffusionAnnePitkäranta,MD,JussiJero,MD,EuricoArruda,MD,AnniVirolainen,MD,andFrederickG.Hayden,MDnoviruses,respiratorysyncytialvirus,Objectives:Tostud
2、ytheassociationofhumanrhinovirus(HRV),respiratoryandhumancoronaviruses.Althoughupsyncytialvirus(RSV),andhumancoronavirusinfectionsinchildrenaged6to80%ofmiddleeareffusionshavebeenmonthsto12yearswithotitismediawitheffusion(OME).Todeterminehowshowntocontainbact
3、erialDNAbylongHRVRNAcanbedetectedafterHRVinfection.polymerasechainreaction–basedampli-ficationtechniques,3,4antibiotictreat-Methods:Middleeareffusion(MEE)samplescollectedatthetimeoftympa-mentinfrequentlyaffectsthecourseofnostomytubeplacementfrom100childrenwi
4、thOMEwereexamined.Viral5,6OME.SeveralearlierstudiesindicateRNAwasdetectedbyreverse-transcriptasepolymerasechainreaction.Forthatvirusesmayplayaroleinthepatho-HRVtheresultswerecomparedwithvirusisolationincellculture.Invitro7,8genesisofOME.Previousdiagnosticstu
5、diesofthepersistenceofHRVinfectivityandRNAwereconductedbycom-toolstodetectHRVinMEEshavebeenbining~105mediancellcultureinfectiousdosesofHRVwithpooledMEEatlimitedtostandardculturemethods,and37°Candassayingserialsamplesfor12weeks.therehasnotbeenapracticaldiagno
6、sticResults:VirusRNAwasdetectedin30children.HRVwasdetectedbyre-methodforHCVinfection.verse-transcriptasepolymerasechainreactionin19childrenwithOMEandbyvirusisolationin5children.RSVRNAwasfoundin8andHCVin3childrenAOMAcuteotitismediaHCVHumancoronaviruswithOME.N
7、odualviralinfectionwasfound.Bacterialpathogenswereisolat-HRVHumanrhinovirusedfrom35MEEsamplesandwereassociatedwithviralRNAin11cases,mostMEEMiddleeareffusionoftenwithHRV(9cases).Underinvitroconditions,HRVculturepositivityde-OMEOtitismediawitheffusionPBSPhosph
8、ate-bufferedsalineclinedrapidly(<2days),butRNAwasdetectableforupto8weeks.RSVRespiratorysyncytialvirusConclusions:Theseresultssuggestthatvirusinfection,particularlyHRVinfec-RT-PCRReverse-transcri