Inventory of Extended-Spectrum-β-Lactamase-Producing Enterobacteriaceae in France as Assessed by a Multicenter Study F. Robin.pdf

该【Inventory of Extended-Spectrum-β-Lactamase-Producing Enterobacteriaceae in France as Assessed by a Multicenter Study F. Robin 】是由【四婆子】上传分享，文档一共【40】页，该文档可以免费在线阅读，需要了解更多关于【Inventory of Extended-Spectrum-β-Lactamase-Producing Enterobacteriaceae in France as Assessed by a Multicenter Study F. Robin 】的内容，可以使用淘豆网的站内搜索功能，选择自己适合的文档，以下文字是截取该文章内的部分文字，如需要获得完整电子版，请下载此文档到您的设备，方便您编辑和打印。:..:-16Copyright?2016,-producingEnterobacteriaceaeinFrance:inventoryassessedbyamulticentricstudy2RobinFa,b,BeyrouthyRa,b,BonacorsiSc,d,AissaNe,BretLf,BrieuNg,CattoirVh,ChapuisAi,3ChardonHg,DegandNj,Doucet-PopulaireFk,DuboisVl,FortineauNm,GrillonAn,LanottePo,Downloadedfrom4LeysseneDp,PatryIq,PodglajenIr,ReculeCs,RosAt,Colomb-CotinatMu,PontiesVu,Ploy5MCv,Ra,-Ferrand,RdelaRésistanceauxAntibiotiques,Clermont-Ferrand,France7bUMRINSERM1071uscINRA2018,Universitéd’Auvergne,M2iSHClermont-Ferrand,Francehttp://aac./8cUMRINSERM1137,IAME,UnivParisDiderot,SorbonneParisCité,F-75018Paris,France9dAP-HP,H?pitalRober-REscherichiacoli,F-75019Paris,France10eLaboratoiredeBactériologie,CHRUdeNancy,Nancy,France11fDepartmentofMicrobiology,LaSourceHospital,CHROrléans,Orléans,France12gCHduPaysd'Aix,LaboratoiredeDiagnosticBiologiquedesMaladiesInfectieusesetd'Hygiène,Aix-en-Provence,France13hCHUdeCaen,ServicedeMicrobiologie,UniversitédeCaenBasse-Normandie,EA4655(équipe"Antibiorésistance"),F-14032Caen,FranceonDecember14,2016byUNIVERSITYOFMEMPHIS14iDepartmentofBacteriology,UniversityHospitalofDijon,BP37013,21070DIJONCEDEX,France15jLaboratoiredeBactériologie,CentreHospitalierUniversitairedeNice,H?pitalArchetII,Nice,France16kServicedeBactériologie-Hygiène,HopitalAntoineBéclère,APHP,UniversitéParisSud,Clamart,FranceRS,UniversityBordeaux2,146rueLeoSaignat,33076Bordeaux,France18mServicedeBactériologie-Virologie,H?pitaldeBicêtre,AssistancePublique/H?pitauxdeParis,94275LeKremlin-Bicêtre,France19nPlateautechniquedemicrobiologie,Laboratoiredebactériologie,CHUStrasbourg,France20oCHUdeTours,LaboratoiredeBactériologie,Tours,France;UniversitéFran?oisRabelais,E5"Bactériesetrisquematerno-f?tal",UMR211282ISP,INRA,Nouzilly,France22pLaboratoiredeBiologiemédicale,CentrehospitalierdelaC?teBasque,64100Bayonne,France23qLaboratoiredeBactériologie,CentreHospitalierUniversitairedeBesan?on,France24rMicrobiology,AP-HP,esPompidou,Paris,France25sBacteriologyDepartment,CHUGrenoble,France26tLaboratoiredeBactériologie-Virologie-Hygiène,CHUdeSaint-Etienne,Saint-Etienne,France27uDepartmentofinfectiousdiseases,InstitutdeVeilleSanitaire,FrenchInstituteforPublicHealthSurveillance,Saint-Maurice,France28vLaboratoiredebactériologie-virologie-hygiène,centrehospitalieruniversitairedeLimoges,87042Limogescedex,France293031Runningtitle:DiversityofESBL,FrenchMulticentricstudy:..32Correspondingauthor:,laboratoiredeBactériologieClinique,CHUClermont-33Ferrand,58,rueMontalembert,Clermont-FerrandF-63003,:33-:33--mail:******@chu-:ESBL,plasmid-mediatedquinoloneresistance,AAC(3)-II,AAC(6’)-Ib,plasmid36http://aac./onDecember14,2016byUNIVERSITYOFMEMPHIS:..37Abstract38OBJECTIVES:TheobjectiveofthisstudywastoperformaninventoryoftheESBL-producing39EnterobacteriaceaeisolatesresponsibleforinfectionsinFrenchhospitals,:200non-redundantESBL-producingEnterobacteriaceaestrainsisolatedfrom42clinicalsampleswerecollectedduringamulti-centricstudyperformedin18representativehttp://aac./-paredbyPCR-based-replicon-typingandplasmid-46multi-locus-sequence-,2016byUNIVERSITYOFMEMPHIS47RESULTS:CTX-M-15,CTX-M-1,CTX-M-14andSHV-12werethemostprevalentESBLs(%).ThethreeCTX-M-typeEBSLsweresignificantlyobservedinEscherichiacoli(%,%%respectively),andCTX-M-15wasthepredominantESBLinKlebsiella50pneumoniae(%).SHV-12wasassociatedwithESBL-encodingEnterobactercloacae51strains(%).qnrB,aac(6’)-Ib-crandaac(3)-IIgeneswerethemainplasmid-mediated52resistancegenes,%-encodingplasmids,,theESBL-:-encodingplasmidsandthebacterialhostexplainthe60differencesobservedintheEnterobacteriaceaespecies.:..616263Downloadedfromhttp://aac./onDecember14,2016byUNIVERSITYOFMEMPHIS:..64Introduction65?-lactamantibiotics,owingtotheirbactericidaleffectandtheirlowtoxicity,,theirincreasingusehasledtotheemergenceofextended68spectrumβ-lactamase(ESBL)-,69cephalosporins,andoxyiminoβ-lactams(ceftazidime,cefotaxime,ceftriaxone,cefepimeandhttp://aac./70aztreonam),butareinactiveagainstcarbapenemsandcephamycins,andaresusceptibleto71clavulanateandtazobactaminhibitionactivity(1).promisedtheuse72ofextendedspectrumcephalosporins,promotedtheuseofcarbapenemsandconsequently73favouredthediffusionofcarbapenemase-,2016byUNIVERSITYOFMEMPHIS74frequentlyresistanttoothermajorclassesofantibioticssuchasaminoglycosidesand75fluoroquinolonesbecauseofassociatedplasmid-’s,TEM-andSHV-(1).Unfortunately,-M-grouphavenowdiffused79worldwide,’s80andnow,theprevalenceofESBL-producingEnterobacteriaceaestrainshasdramatically81increasedinmanycountries,,becauseofthediffusionofCTX-82Menzymes(1,2).83AlthoughESBLshavebeenwidelyinvestigated,therehavebeentodatenomulticentric84studiesofESBL-,wecollected85200ESBL--:..://aac./onDecember14,2016byUNIVERSITYOFMEMPHIS:..90MaterialandMethods91ClinicalisolatesDownloadedfrom92BetweenJune2012andOctober2012,consecutive,non-repetitiveESBL-producing93Enterobacteriaceaestrainsisolatedfromclinicalsamplesofinpatientsandoutpatientswere94collectedin18representativeFrenchhospitals(15teachinghospitalsand3non-teaching95publichospitals)://aac./96wereincludedandESBL-ordingtomitteeoftheFrenchSocietyfor98Microbiology(CA-SFM)(3).Screeningsamples,suchasrectalswabs,,(GabrielMontpiedHospital,Clermont-Ferrand).(Robert-Debréhospital,Paris).104AntimicrobialsusceptibilitytestingandESBLdetection105IdentificationofstrainswasconfirmedbytheVitekMSMALDITOFmethod(bioMérieux,La106Balme,France).mendationsofmitteeoftheFrenchSocietyforMicrobiology(CA-SFM)(3).AntibioticordingtotheCA-SFMmendations(3).ESBL-productionwasconfirmedbythedouble-disksynergytest(3).11018followingantibioticsweretested:amoxicillin,amoxicillin-clavulanicacid,ticarcillin,111piperacillin-tazobactam,cephalothin,cefoxitin,ceftazidime,cefotaxime,ertapenem,112imipenem,amikacin,gentamicin,tobramycin,nalidixicacid,ofloxacin,ciprofloxacin,113fosfomycinandcotrimoxazole(Bio-Rad,Marnes-La-Coquette,France).:..114Characterizationofβ-lactamase-encodinggenesandassociatedresistancegenes115β-lactamaseswereinvestigatedbyisoelectricfocusingaspreviouslydescribed(4).TheESBL-116encodinggeneswereidentifiedbyspecificPCRandsequencingexperimentsusingspecificDownloadedfrom117primersasdescribedelsewhere(4).Carbapenemaseproductionwasinvestigatedin118carbapenem-resistantstrainsbyspecificPCRaspreviouslydescribed(5).Genesencoding119AAC(6’)-Ib,AAC(6’)-Ib-cr,AAC(3)-II,RmtAtoH,ArmA,NpmA,Qnr(A,B,C,D,S),Oqx(forhttp://aac./120strainsnotbelongingtoKlebsiellagenus)andQepAwereidentifiedbyspecificPCRassayand121sequencingaspreviouslyreported(6–16).Plasmid-mediatedcolistinresistancemechanism122MCR-1wasinvestigatedaspreviouslydescribed(17).Plasmid-encodedAmpCandOXA-1123productionwereinvestigatedaspreviouslydescribed(18,19).onDecember14,2016byUNIVERSITYOFMEMPHIS124Molecularstraintyping125TherelatednessbetweenESBL-producingisolateswasassessedbyrepetitivesequence-126basedPCR(rep-PCR)ordingtothemanufacturer's127instructions(bioMérieux,LaBalme,France).TheanalysiswasperformedusingPearson's128correlationinthededicatedDLsoftwareofthemanufacturer().Isolateswith129similaritiesof<95%wereconsidereddifferent,andisolateswithsimilaritiesof>98%>95%and<98%werejudged131manuallyusingthepatternoverlayoftheanalysistoolinthesoftware(20).APearson132distancematrixwascalculatedusingDiversilabandaclassicalmultidimensionalscalingwas133thenperformedonthedistancematrixusingRtoillustratetherelationshipbetweenthe134isolates(21).(A,B1,B2,C,D,E,F)wasdeterminedbya135PCRmethodaspreviouslydescribed(22).ThehighlyvirulentB21subgroupwasalso136identifiedbyspecificPCR(23).O25b-:..137PCRmethoddescribedbyClermontetal.(24).(25).Downloadedfrom139AnalysisofESBL--encodinggeneswasstudiedbymating-,plasmidDNAwasextractedwithNucleoBond?Xtra(Macherey-Nagel,142Germany)://aac./143performedonagarplatessupplementedwithceftazidimeorcefotaxime(4mg/L)and144rifampicin(300mg/L)forthemating-outassayandceftazidimeorcefotaxime(4mg/L)-encodingplasmidswasdeterminedbySouthernblotwith146ESBLspecificprobesusingplasmidDNAextractedbythemethodofKadoandLiu(26)withonDecember14,2016byUNIVERSITYOFMEMPHIS147theplasmidsRsa(39kb),TP114(61kb),pCFF04(85kb)andpCFF14(180kb)-basedreplicontyping(PBRT)andplasmidmultilocussequence150typing(27,28).151Characterizationofvirulencefactor-encodinggenes152The11followingvirulencefactor-encodinggenesofextra-(ExPEC)wereidentifiedbymultiplexPCR,,aspreviouslydescribed(29):papC(outer154membraneusherproteinPAPC),papGIIandpapGIII(Pfimbriae),sfa/foc(Sfimbriae),155hlyC(hemolysin),cnf1(ecrotizingfactor),(aerobactin),fyuA(156yersiniabactin),iroN(salmochelin),hek/hra(haemagglutinin)andibeA(endothelial157invasin).,9genes(uge,mrkD,fimH,allS,rmpA,magA,wabG,kfu,158cf29a)werescreenedbyPCRassays(30).Capsulecsp-basedgenotypingwasperformedas:..159previouslydescribed(31).StrainsNTUH-K2044,://aac./onDecember14,2016byUNIVERSITYOFMEMPHIS:..162ResultsontextDownloadedfrom164MostESBL-(62%),(%)(%)species(Figure1a).Annual166prevalencewascalc