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1、检验医学2011年1月第26卷第1期LaboratoryMedicine,January2011,Vol26.No1.51文章编号:16738640(2011)01005105中图分类号:R378.2文献标识码:A大肠埃希菌和肺炎克雷伯菌外排泵转录水平与耐药的关系蒋燕群,李俐,钱燕斐(上海交通大学附属第六人民医院检验科,上海200233)摘要:目的调查细菌耐药性与其主动外排泵系统的作用关系,并探讨大肠埃希菌外排泵调控基因对其外排泵结构基因表达水平的影响,分析其耐药机制。方法收集大肠埃希菌和肺炎克雷伯菌共59株,根据头孢他啶耐药情况分为实验组(耐药)与对照组
2、(敏感),多重聚合酶链反应(PCR)扩增外排泵调控基因acrR并测序,实时定量逆转录(RT)PCR检测外排泵及调控基因marA的相对转表达水平,三维试验检测AmpC酶。结果实验组大肠埃希菌46.7%(7株)acrAB基因转录水平增高,肺炎克雷伯菌中51.9%(14株)acrAkp基因转录水平增高,敏感组外排泵基因均未见增高。实验组大肠埃希菌中,3株acrR发现突变,9株marA基因转录水平增高。59株菌株中39株(66.1%)AmpC阳性。结论主动外排泵系统是细菌耐药机制之一,常与其他耐药机制共同存在。acrR突变与marA表达增高均可引起大肠埃希菌acrAB表达增加。关键词:大肠埃
3、希菌;肺炎克雷伯菌;外排泵;耐药RelationshipbetweenthelevelofeffluxpumpinEscherichiacoliandKlebsiellapneumoniaanddrugresistanceJIANGYanqun,LILi,QIANYanfei.(DepartmentofClinicalLaboratory,theSixthPeoplesHospital,ShanghaiJiaotongUniversitySchoolofMedicine,Shanghai200233,China)Abstract:ObjectiveToinvestigate
4、therelationshipbetweenbacterialresistanceanditsactiveroleofeffluxpumpsystems,exploretheinfluencebetweentheregulatorygenesofeffluxpumpinEscherichiacoliandtheexpressionlevelofitsstructuregene,andanalyzetheresistantmechanism.Methods59strainsofEscherichiacoliandKlebsiellapneumoniawerecollected,andthe
5、strainswereclassifiedintotwogroupsasexperimentalgroupandcontrolgroupaccordingtotheresistanceofceftazidime.Themultiplexpolymerasechainreaction(PCR)wasusedfortheamplificationandseguencingoftheregulatorygeneacrR,andtherealtime(RT)PCRwasusedforthedetectionoftheexpressionlevelofeffluxpumpandtheregulat
6、orygenemarA.ThethreedimensiontestwasusedforthedetectionofAmpCbetalactamase.Results7strainsofEscherichiacoli(46.7%)hadhighacrABtranscriptionlevelinexperimentalgroup,and14strainsofKlebsiellapneumonia(51.9%)hadhighacrAkptranscriptionlevelinexperimentalgroup.AmongtheEscherichiacoliinexperimentalgro
7、up,3isolateshadmutationofacrRgene,and9isolateshadhighmarAgenetranscriptionleve.lAmpCbetalactamasewasdetectedin39of59(66.1%)isolates.ConclusionsTheeffluxpumpsystemisoneofthemechanismsofbacterialresistance,whicho
