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Detection of NDM-1-Producing Klebsiella pneumoniae in Kenya

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Published Ahead of Print 29 November 2010. 10.1128/AAC.01247-10. 2011, 55(2):934. DOI:Antimicrob. Agents Chemother. Patrice NordmannLaurent Poirel, Gunturu Revathi, Sandrine Bernabeu and in Kenya pneumoniaeKlebsiellaDetection of NDM-1-Producing http://aac.asm.org/content/55/2/934Updated information and services can be found at: These include:REFERENCEShttp://aac.asm.org/content/55/2/934#ref-list-1at: This article cites 18 articles, 10 of which can be accessed free

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DetectionofNDM-1-ProducingKlebsiellapneumoniaeinKenyaᰔLaurentPoirel,1GunturuRevathi,2SandrineBernabeu,1andPatriceNordmann1*ServicedeBacte´riologie-Virologie,INSERMU914EmergingResistancetoAntibiotics,HoˆpitaldeBiceˆtre,AssistancePublique/HoˆpitauxdeParis,Faculte´deMe´decineParisSud,K.-Biceˆtre,France,1andDepartmentofPathology,TheAgaKhanUniversityHospital,Nairobi,Kenya2

Received9September2010/Returnedformodification24October2010/Accepted21November2010Sevencarbapenem-resistantNDM-1-positiveKlebsiellapneumoniaeisolateswererecoveredfrompatientshospitalizedbetween2007and2009indifferentwardsatareferralandtertiarycarecenterinNairobi.Mostoftheisolateswereobtainedfromurine.AllisolatescarriedtheblaNDM-1carbapenemasegenepreviously

reportedfromIndia,Pakistan,andtheUnitedKingdom.Theseisolateswereclonallyrelatedandexpressedmanyotherresistancedeterminants,including␤-lactamasesCTX-M-15,OXA-1,OXA-9,CMY-6,andamino-glycosideresistancemethylaseRmtC.ThisworkcorrespondstothefirstreportofNDM-1producersinAfrica.

Carbapenem-hydrolyzing␤-lactamasesbelongingtoAmblerclassesA,B,andDhavebeenreportedworldwideamongEnterobacteriaceae(10,14,18).ThemostclinicallysignificantareKPC-type(AmblerclassA),IMPandVIMtypes(classB),andOXA-48(classD),mostlyidentifiedinKlebsiellapneu-moniaeasasourceofnosocomialoutbreaks(14).Mostofthoseisolatesaremultidrugresistant.Recently,anovelmetallo-␤-lactamase(MBL)namedNDM-1(NewDelhimetallo-␤-lactamase)hasbeenidentifiedfromK.pneumoniae(strain05-506)andEscherichiacoliiso-latesrecoveredinSwedenfromapatientpreviouslyhospital-izedinIndia(19).AnextensivesurveyperformedintheUnitedKingdom,India,andPakistanidentifiedNDM-1-pro-ducingK.pneumoniae,E.coli,Citrobacterfreundii,Morganellamorganii,Providenciaspp.,andEnterobactercloacaeisolates(9).ThepresenceofNDM-1producersinhospitalizedpatientsintheUnitedKingdomwasrelatedinmanycasestoprevioushospitalizationintheIndiansubcontinent(9),andsimilarob-servationshavebeenmadeinFrance(12,15)andtheSultan-ateofOman(11).SpreadofthisnovelcarbapenemasegeneisconsideredaseriousthreatsincethereservoirofNDM-1pro-ducersisatleastinpartrelatedtotheIndiansubcontinent,whichisinhabitedbythesecond-largestpopulationintheworldandwhereNDM-1producersarereportedalsoincom-munity-acquiredinfections(9,12).Ourstudywasinitiatedusinganarchivalcollectionofbac-terialisolatesfromtheAgaKhanUniversityHospital,Nairobi,Kenya,forthepurposeofsurveillanceofantibacterialresis-tancemechanismsandinfectioncontrolaudits.Sevenmulti-drug-resistantK.pneumoniaestrainsisolatedovera3-yearperiod(2007to2009)wereselected,allshowinganidenticalpatternofresistance.Thoseisolateshadbeenrecoveredmostlyfromurine,frompatientswhowereallreceivinganti-biotictreatmentswhenthesampleshadbeenrecovered(Table1).However,thosetreatmentswerenottargetedtowardthe

carbapenem-resistantK.pneumoniaeisolatesthatweresystem-aticallyconsiderednon-strictlyinvasive(Table1).TheMICsweredeterminedbyEtest(ABbioMe´rieux,Solna,Sweden)onMueller-Hintonagarplatesandbyliquidmicrodilutionassaysat37°C,andresultsofsusceptibilitytest-ingwereinterpretedaccordingtotheCLSIguidelines(6).Theisolateswereresistanttoall␤-lactams,includingcarbapenemswithMICvaluesofϾ32␮g/mlforimipenem,meropenem,ertapenem,anddoripenem.Inaddition,theywereresistanttoaminoglycosides,fluoroquinolones,chloramphenicol,sulfon-amides,fosfomycin,andnitrofurantoin,theMICofrifampinwasϾ32␮g/ml,andtheMICsoftigecyclineandcolistinmea-suredbyEtestwere0.5and0.5␮g/ml,respectively.ResultsfromMBLdetectionperformedbyusingEtestMBLstrips(ABbioMe´rieux)werepositive.Thus,PCRassayswerecarriedoutwithaseriesofprimersdesignedforthedetectionofseveralclassB␤-lactamasegenes,blaIMP,blaVIM(16),and

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新型NDM-1酶抑制剂的筛选与研究

新型NDM-1酶抑制剂的筛选与研究

新型NDM-1酶抑制剂的筛选与研究王颖;刘忆霜;李兴华;白银磊;鲁众阳;肖春玲【摘要】目的建立肺炎克雷伯菌(Klebsiella pneumoniae)新德里β-内酰胺酶1(New Delhi metallo-β-lactamase-1,NDM-1)抑制剂的高通量筛选模型,从不同来源的样品中筛选该酶抑制剂,以期获得作用于NDM-1的新型抗耐药阴性菌的药物或药物先导物.方法克隆、表达并纯化肺炎克雷伯菌ATCC BAA-2146的NDM-1酶;基于在NDM-1作用下头孢硝噻吩溶液由黄色变成红色,且在486nm波长下具有光吸收的原理,建立酶活测定体系,构建和评价NDM-1抑制剂的高通量筛选模型;应用此模型筛选NDM-1抑制剂,评价其作为抗耐药阴性菌药物的可行性.结果得到了重组NDM-1酶,构建了NDM-1酶抑制剂高通量筛选模型,筛选得到5个NDM-1抑制剂,其中化合物IMB-XW2为该酶的非竞争性抑制剂,其Ki值为4.6μmol/L,IC50为10.3 μmol/L;该化合物在4μg/mL时可以使表达NDM-1的大肠埃希菌对美罗培南的敏感度提高8倍.结论成功构建了稳定性好、灵敏度高的NDM-1酶抑制剂的高通量筛选模型,应用该模型可筛选到NDM-1抑制剂,该抑制剂有可能成为新型的抗耐药革兰阴性菌的药物或药物先导物.【期刊名称】《中国抗生素杂志》【年(卷),期】2015(040)006【总页数】6页(P401-406)【关键词】NDM-1;高通量筛选;酶抑制剂【作者】王颖;刘忆霜;李兴华;白银磊;鲁众阳;肖春玲【作者单位】北京协和医学院中国医学科学院医药生物技术研究所,北京100050;北京协和医学院中国医学科学院医药生物技术研究所,北京100050;北京协和医学院中国医学科学院医药生物技术研究所,北京100050;北京协和医学院中国医学科学院医药生物技术研究所,北京100050;北京协和医学院中国医学科学院医药生物技术研究所,北京100050;北京协和医学院中国医学科学院医药生物技术研究所,北京100050【正文语种】中文【中图分类】R978.1随着抗生素的长期和广泛使用，细菌耐药问题日趋严重，致病菌中多药耐药菌株检出率的不断增加，耐药细菌感染的临床治疗面临巨大挑战。

耐碳青霉烯类肺炎克雷伯菌耐药机制研究

耐碳青霉烯类肺炎克雷伯菌耐药机制研究

耐碳青霉烯类肺炎克雷伯菌耐药机制研究黄峰;许元元;秦淑国【摘要】目的:探讨耐碳青霉烯类肺炎克雷伯菌耐药基因及耐药特性.方法:对42株耐碳青霉烯类肺炎克雷伯菌进行药敏试验、改良Hodge试验,并采用聚合酶链反应(PCR)方法检测细菌肺炎克雷伯菌碳青霉烯酶(KPC)基因.结果:42株耐碳青霉烯类肺炎克雷伯菌中,药敏试验结果显示耐药情况严重,仅对替加环素、阿米卡星、妥布霉素、复方新诺明较敏感.改良Hodge试验及KPC基因检测全部阳性,基因测序为KPC-2型.结论:KPC-2是引起肺炎克雷伯菌耐药的主要原因,应引起临床及实验室的关注.%Objective:To explore the drug resistance gene of Klebsiella pneumoniae carbapenemase,and its resistant features.Methods:The drug sensitivity test and modified Hodge test in 42 strains of Klebsiella pneumoniae carbapenemase were performed,and the KPC gene of bacterium was detected using PCR.Results:The drug resistance in 42 strains of Klebsiella pneumoniae carbapenemase were severe,which was sensitive to the tigecycline,amikacin,tobramycin and cotrimoxazole.The detection results of modified Hodge test and KPC gene were positive,and the type KPC-2 gene was identified.Conclusions:KPC-2 is the major cause of drug resistance of Klebsiella pneumoniae,which should be paid attention to in clinic and laboratory.【期刊名称】《蚌埠医学院学报》【年(卷),期】2017(042)003【总页数】4页(P379-382)【关键词】耐碳青霉烯类肺炎克雷伯菌;耐药机制;改良Hodge试验【作者】黄峰;许元元;秦淑国【作者单位】皖北煤电集团总医院(蚌埠医学院第三附属医院) 检验科,安徽宿州234000;皖北煤电集团总医院(蚌埠医学院第三附属医院) 检验科,安徽宿州234000;皖北煤电集团总医院(蚌埠医学院第三附属医院) 检验科,安徽宿州234000【正文语种】中文【中图分类】R378.99肠杆菌科细菌作为医院感染和社区获得性感染的重要病原菌，可引起呼吸道、尿路等各种部位的感染以及菌血症，近年来由于抗菌药物的大量及不合理使用促进了耐药菌株的迅速扩散。

猪源肺炎克雷伯氏菌耐药性及毒力基因分析

猪源肺炎克雷伯氏菌耐药性及毒力基因分析

102猪源肺炎克雷伯氏菌耐药性及毒力基因分析欧冠标1，周雨晴1，彭　昊2，陆泽宁3，廖玉英2，马东鑫1，袁敬知1，葛　强1，李　珣1，王晓晔1（1.广西大学动物科学技术学院，广西南宁　530004；2. 广西壮族自治区兽医研究所，广西南宁　530000；3.南宁市武鸣区双桥镇水产畜牧兽医站，广西南宁　530104）摘　要：为了解猪群中猪源肺炎克雷伯氏菌耐药情况和毒力基因携带状况，2019年采集存栏生猪肛拭子样品125份、宰后生猪肉类样品50份，通过细菌分离培养、药敏试验以及PCR 扩增16S rRNA 、khe 基因、耐药基因和毒力基因等方法对肺炎克雷伯氏菌特性进行分析。

结果显示，从125份肛棉拭子样品、50份宰后生猪肉类样品中各分离得到32、4株肺炎克雷伯氏菌；药敏试验结果显示，分离的肺炎克雷伯氏菌对四环素耐药性最高（83.3%），其次是氯霉素（63.9%）。

对36株分离菌进行耐药基因和毒力基因检测，发现携带耐药基因tet(A)的达30株、携带floR 、mcr-1的分别为23、10株，且有23株菌同时携带tet(A)和floR 基因，少部份菌株携带aadA1、bla TEM 、bla CTX-M-1、bla OXA 、qnrB 、aac(6')-Ib -cr ；携带毒力基因wabG 的达26株、携带uge 和fimH 的25株、携带kfu 的23株、携带aereobactin 的7株，且有20株菌同时携带wabG 、uge 、fimH 和kfu 基因。

结果表明，我国猪群中肺炎克雷伯氏菌检出率可能较高，菌株携带耐药基因及毒力基因较为集中，对部分抗生素耐药性较强。

本研究为肺炎克雷伯氏菌的检测、诊断以及合理使用抗生素治疗提供了试验依据，同时也为猪源肺炎克雷伯氏菌防控提供了数据支撑。

关键词：猪源肺炎克雷伯氏菌；PCR 鉴定；药敏试验；耐药基因；毒力基因中图分类号：S852.61 文献标识码：A 文章编号：1005-944X （2021）06-0102-10DOI ：10.3969/j.issn.1005-944X.2021.06.021 开放科学（资源服务）标识码（OSID ）：Analysis on Drug Resistance and Virulence Genes of Porcine Klebsiella pneumoniaeOu Guanbiao 1，Zhou Yuqing 1，Peng Hao 2，Lu Zening 3，Liao Yuying 2，Ma Dongxin 1，Yuan Jingzhi 1，Ge Qiang 1，Li Xun 1，Wang Xiaoye 1（1. College of Animal Science and Technology ，Guangxi University ，Nanning ，Guangxi 530004，China ；2. Veterinary Research Institute of Guangxi Zhuang Autonomous Region ，Nanjing ，Guangxi 530000，China ；3. Fisheries ，Animal Husbandry and V eterinary Station of Shuangqiao Township ，Nanning ，Guangxi 530104，China ）Abstract ：In 2019，in order to investigate the status of drug resistance and carrying virulence genes of porcine Klebsiella pneumoniae （K. pneumoniae ），125 anal swabs and 50 fresh pork samples were collected respectively from live and slaughtered pigs to analyze the characteristics of K. pneumoniae through isolation and culture of bacteria ，drug sensitivity test and amplification of 16S rRNA and the Khe ，drug resistance and virulence genes by PCR. The results showed that ，32 and 4 strains of K. pneumoniae were isolated respectively from anal swabs and fresh pork samples. 收稿日期：2021-02-07　修回日期：2021-04-16基金项目：国家现代农业产业体系广西肉鸡产业创新团队建设专项（nycytxgxcxtd ）；南宁市科学研究与技术 开发计划项目（20202106）；崇左市科技计划项目（崇科FA 2019006）；南宁市武鸣区科学研究与 技术开发项目（20190101）同等贡献作者：欧冠标，周雨晴通信作者：王晓晔。

耐碳青霉烯肺炎克雷伯菌感染的危险因素及耐药机制的研究

耐碳青霉烯肺炎克雷伯菌感染的危险因素及耐药机制的研究

耐碳青霉烯肺炎克雷伯菌感染的危险因素及耐药机制的研究胡小飞厉银平刘桂霞彭清臻王锋【摘要】目的探讨耐碳青霉烯肺炎克雷伯菌感染的危险因素及耐药机制。

方法对我院2018年6月1日-2019年5月31日57例耐碳青霉烯肺炎克雷伯菌(CRKP)感染患者和70例碳青霉烯敏感肺炎克雷伯菌(CSKP)感染患者进行回顾性分析。

结果CRKP感染患者主要分布在呼吸科31.6%(18/57)、神经内科21.1%(12/57)、神经外科14%(8/57)和ILU10.5%(6/57)，感染部位以下呼吸道感染78.9%(45/57)和泌尿道感染17.8%(8/57)为主;相比CSKP感染组,使用两种或三种抗生素患者的比例在CRKP感染组中显著增加。

多元L wp U c回归分析显示,1月内使用三代头抱/酶抑制剂、1月内使用碳青霉烯类以及留置尿管为CRKP感染的独立危险因素。

57株CRKP菌株中，有39株(68.4%)携带biKPC基因,6株(10.5%)携带bilMP基因$结论CRKP菌株日见增多，规范化使用碳青霉烯类抗生素，减少或缩短侵入性医疗操作有利于降低CRKP感染的发生率;产KPC型碳青霉烯酶是CRKP菌株对碳青酶烯类耐药的最主要耐药机制。

【关键词】肺炎克雷伯菌;碳青霉烯类耐药;危险因素;碳青霉烯酶基因Risk factors and drrg resistancc mechanism of carbapenem-resistant KUbsieea pnenmoniae infectionHU Xiao-fei,LI Yitfing,LI Gui-fi,PENG Qing-zZee,WANG FengDepartment of RespiratorS and Critical Can Medicine,Xiaogan Hospital0ffiliate0m Wuhan Universito f Science and TecCnology,Xiaogan,Hubei432000,China(Abstract]Objective To investigate the risk factors and drug resistance mechanism of carbapenem-resistant Klebsiella pneumoniae infection.Methods A retrospective analysis was conducted on57patients with carbapenem resistant Klebsiella pneumoniae(CRKP)infection and70patienO with carbapenem sensitive Klebsiella pneumoniae (CSKP)infection in our hospital from June1,2018to May31,2019.ResUs The patients with CRKP infection weeemainiydisieibuied in eespieaioeydepaeimeni(31.6%),neueoiogydepaeimeni(21.1%),neueosuegeeydepaei-meni(14%)and ICU(10.5%).Loweeeespieaioeyieaciineeciion(78.9%)and ueinaeyieaciineeciion(17.8%) weeeihemain ineeciion paeed wiih iheCSKPineeciion geoup,ihepeopoeiion oepaiienisusingiwooeiheee aniibioiicsinceeased signieicaniiyin iheCRKPineeciion geoup.Muiiipieiogisiiceegee s ion anaiysisshowed ihaiihe use of thPd-aeneration cephalosporins/enzyme inhibitors within one month,carbapenem antibjotics within one month and indwe i ingcaiheieeweeeindependenieik eacioeeoeCRKPineeciion.Oeihe57CRKPHieainH,39(68.4%)cae-ried blaKPC gene and6(10.5%)carried blalMP gene.Conclusion The number of CRKP strains is increasing day by day,standardizing the use of carbapenem antibiotics and reducing or shortening invesive medical procedures are beneeiciaiioeeduceiheincidenceoeCRKPineeciion.Thepeoduciion oeKPCiypecaebapenem isihemosiimpoeiani mechanism oecaebapenem eesisianceoeCRKPsieains.(Keywords]Klebsiella pneumoniae;carbapenem-resistant;risk factor;carbapenemase gene肺炎克雷伯菌是医院获得性感染检出排名仅次于大肠埃希菌的革兰氏阴性杆菌,是造成呼吸道、泌尿道、血流等多个部位感染的重要致病菌之一。