PCR-Based Identification of Klebsiella pneumoniae subsp. rhinoscleromatis, the Agent of Rhinoscleroma 英文参考文献.docVIP

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PCR-Based Identification of Klebsiella pneumoniae subsp. rhinoscleromatis, the Agent of Rhinoscleroma 英文参考文献.doc

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PCR-Based Identification of Klebsiella pneumoniae subsp. rhinoscleromatis, the Agent of Rhinoscleroma 英文参考文献

PCR-BasedIdentificationofKlebsiellapneumoniae subsp.rhinoscleromatis,theAgentofRhinoscleroma CindyFevre1,VirginiePasset1,AlexisDeletoile1,Vale′rieBarbe2,LionelFrangeul3,AnaS.Almeida4,5, PhilippeSansonetti4,5,Re′gisTournebize4,5,SylvainBrisse1* 1Institut Pasteur, GenotypingofPathogens andPublicHealth,Paris,France, 2CEA-IG,Genoscope,Evry, France, 3InstitutPasteur,Inte′grationet Analyse Ge′nomique, Paris,France,4InstitutPasteur,Unite′ dePathoge′nie MicrobienneMole′culaire,Paris,France,5Unite′ INSERMU786,InstitutPasteur,Paris,France Abstract RhinoscleromaisachronicgranulomatousinfectionoftheupperairwayscausedbythebacteriumKlebsiellapneumoniae subsp. rhinoscleromatis.The disease is endemicin tropical andsubtropical areas, but its diagnosis remains difficult. As a consequence,anddespiteavailableantibiotherapy,somepatientsevolveadvancedstagesthatcanleadtodisfiguration, severerespiratoryimpairmentanddeathbyanoxia.Becauseidentificationoftheetiologicagentiscrucialforthedefinitive diagnosisofthedisease,theaimofthisstudywastodeveloptwosimplePCRassays.Wetookadvantageofthefactthatall Klebsiellapneumoniaesubsp.rhinoscleromatisisolatesare(i)ofcapsularserotypeK3;and(ii)belongtoasingleclonewith diagnosticsinglenucleotidepolymorphisms(SNP).Thecompletesequenceofthegenomicregioncomprisingthecapsular polysaccharidesynthesis(cps)geneclusterwasdetermined.Putativefunctionsofthe21genesidentifiedwereconsistent withthestructureoftheK3antigen.TheK3-specificsequenceofgeneKr11509(wzy)wasexploitedtosetupaPCRtest, which was positive for 40 K3 strains but negative when assayed on the 76 other Klebsiella capsular types. Further, to discriminate Klebsiella pneumoniae subsp. rhinoscleromatis from other K3 Klebsiella strains, a specific PCR assay was developedbasedondiagnosticSNPsinthephosphateporingene phoE.Thisworkprovidesrapidandsimplemolecular tools to confirm the diagnostic of rhinoscleroma, which should improve patient care as well as knowledge on the prevalenceandepidemiologyofrh