碳青霉烯类耐药肠杆菌目细菌耐药性及耐药基因分析  

Analysis on drug resistance and drug resistance genes of carbapenem-resistant Enterobacteriaceae bacteria

作　　者：侯素君 尹盟盟 王均梅[1] 别海文 梁永娟 朱元祺[2] HOU Sujun;YIN Mengmeng;WANG Junmei;BIE Haiwen;LIANG Yongjuan;ZHU Yuanqi(Department of Clinical Laboratory,Rizhao Municipal Hospital of Traditional Chinese Medicine,Rizhao,Shandong 276800,China;Department of Clinical Laboratory,Affiliated Hospital of Qingdao University,Qingdao,Shandong 266003,China)

机构地区：[1]山东省日照市中医医院检验科,山东日照276800 [2]青岛大学附属医院检验科,山东青岛266003

出　　处：《检验医学与临床》2023年第18期2667-2671,共5页Laboratory Medicine and Clinic

基　　金：青岛大学附属医院“临床医学+X”科研项目(2019-3399);山东省日照市中医医院2021年度院级科研立项项目(RZZY2021LC-09)。

摘　　要：目的探讨医院碳青霉烯类耐药肠杆菌目细菌(CRE)的临床耐药性及耐药表型和基因型,为医院CRE菌株感染的临床治疗及医院感染暴发的预防和控制提供科学依据。方法收集2020年7月至2023年1月日照市中医医院临床患者标本中分离的77株CRE菌株,采用全自动微生物分析仪进行细菌鉴定和药敏试验,采用改良碳青霉烯酶灭活试验(mCIM)和乙二胺四乙酸(EDTA)改良碳青霉烯酶灭活试验(eCIM)联合检测碳青霉烯酶表型,采用聚合酶链反应(PCR)扩增耐药基因并进行测序分型。结果77株CRE菌株主要分离自痰液,其次是尿液、胸腔积液;科室来源主要是颅脑外科、重症医学科、脑卒中监护室。CRE菌株中肺炎克雷伯菌33株,大肠埃希菌15株,阴沟肠杆菌12株,其他细菌17株。CRE菌株对替加环素敏感性为97.4%,阿米卡星为44.2%,复方磺胺甲噁唑为39.0%,对其他抗菌药物的敏感性均低于20.0%。mCIM筛选碳青霉烯酶菌株的灵敏度为95.9%,特异度为100.0%。PCR扩增结果显示49株CRE菌株携带NDM基因,24株CRE菌株携带KPC基因,1株CRE菌株携带IMP基因,3株未检出碳青霉烯酶基因;基因测序比对确定耐药基因亚型分别为bla KPC-2、bla NDM-1、bla IMP-4。结论CRE菌株以肺炎克雷伯菌、大肠埃希菌、阴沟肠杆菌为主,具有较强的耐药性,肺炎克雷伯菌碳青霉烯类耐药基因主要为bla KPC-2、大肠埃希菌耐药基因主要为bla NDM-1,阴沟肠杆菌耐药基因主要为bla NDM-1,临床需加强CRE菌株检测,根据药敏试验结果合理选用抗菌药物,防范其在医院内暴发流行。Objective To explore the clinical drug resistance,drug resistance phenotypes and genotypes of carbapenem-resistant Enterobacteriaceae(CRE)bacteria in the hospital so as to provide a scientific basis for the clinical treatment of CRE bacterial strains infection and the prevention and control of the CRE bacterial strains infection outbreaks.Methods Seventy-seven CRE strains isolated from the clinical patients samples in Rizhao Municipal Hospital of Traditional Chinese Medicine from July 2020 to January 2023 were collected,the bacterial identification and drug susceptibility tests were conducted by the full automatic microbiological analyzer,and the carbapenase phenotype was detected by the modified carbapenenase inactivation method(mCIM)and EDTA modified carbapenenase inactivation method(eCIM).The drug-resistance genes were amplified by polymerase chain reaction(PCR)and the sequencing was performed.Results Seventy-seven CRE strains were mainly isolated from the sputum specimen,followed by the urine and pleural effusion.The departments were mainly the craniocerebral surgery,intensive care medicine and stroke intensive care unit.Among the CRE strains,there were 33 strains of Klebsiella pneumoniae,15 strains of Escherichia coli,12 strains of Enterobacter cloacae and 17 strains of other bacteria.The sensitivity of the CRE strains to tigacycline was 97.4%,which to amikacin was 44.2%,which to cotrimoxazole was 39.0%,and which to other antibacterial drugs was lower than 20.0%.The sensitivity and specificity of mCTM for screening carbapenase strain were 95.9%and 100.0%,respectively.The PCR amplification results showed that 49 strains of CRE bacteria carried the NDM gene,24 strains of CRE bacteria carried KPC gene,1 strain of CRE bacterium carried IMP gene,and 3 strains did not detect carbapenenase gene.The gene sequencing comparison determined that the subtypes of drug resistance gene were bla KPC-2,bla NDM-1 and bla IMP-4.Conclusion The CRE strains are mainly Klebsiella pneumoniae,Escherichia coli and Enterobacter cloacae

关 键 词：碳青霉烯类耐药肠杆菌目细菌 细菌耐药性 耐药基因 

分 类 号：R446.5[医药卫生—诊断学]