CAJ | 학술논문

目的分析我国对碳青霉烯类药物敏感性降低肠杆菌科细菌中碳青霉烯酶基因的携带情况及分子流行病学特征.方法收集全国1 1个城市14家医院临床分离的碳青霉烯类药物不敏感肠杆菌科细菌201株,采用琼脂稀释法检测14种抗菌药物的MICs,改良Hodge试验和乙二胺四乙酸(EDTA)协同试验筛检碳青霉烯酶表型,PCR扩增测序确证菌株携带的耐药基因.对产碳青霉烯酶菌株进行接合试验,脉冲场凝胶电泳(PFGE)和多位点序列分型(MLST)分析.结果 201株碳青霉烯类药物敏感性降低肠杆菌科细菌中有53株确定为产碳青霉烯酶[KPC-2(32株)、IMP-4(20株)、IMP-8(1株)],其中肺炎克雷伯菌33株、弗劳地枸橼酸杆菌9株、大肠埃希菌6株、阴沟肠杆菌5株,分别来自于北京(43株)、杭州(6株)、南京(3株)、福州(1株)4个地区;将53株产酶菌株与大肠埃希菌EC600进行接合,获得28株接合子;PFGE分型结果显示,肺炎克雷伯菌(33株)可分为3个型2个亚型、弗劳地枸橼酸杆菌(9株)可分为4个型、阴沟肠杆菌(5株)可分为4个型、大肠埃希菌(6株)均为同一型;29株产KPC-2型酶肺炎克雷伯菌的MLST分型序列均为ST11,4株产IMP-4型酶肺炎克雷伯菌中3株为ST876,1株为ST147.结论在对我国11个城市的耐药菌监测中只有北京、杭州、南京和福州的医院中检出携带碳青霉烯酶基因的菌株,其中KPC-2最常见,其次为IMP-4、IMP-8,菌株间同源性较高,由于碳青霉烯酶基因在菌株问容易发生水平转移,应引起临床高度关注.
목적 분석아국대탄청매희류약물민감성강저장간균과세균중탄청매희매기인적휴대정황급분자류행병학특정.방법 수집전국1 1개성시14가의원림상분리적탄청매희류약물불민감장간균과세균201주,채용경지희석법검측14충항균약물적MICs,개량Hodge시험화을이알사을산(EDTA)협동시험사검탄청매희매표형,PCR확증측서학증균주휴대적내약기인.대산탄청매희매균주진행접합시험,맥충장응효전영(PFGE)화다위점서렬분형(MLST)분석.결과 201주탄청매희류약물민감성강저장간균과세균중유53주학정위산탄청매희매[KPC-2(32주)、IMP-4(20주)、IMP-8(1주)],기중폐염극뢰백균33주、불로지구연산간균9주、대장애희균6주、음구장간균5주,분별래자우북경(43주)、항주(6주)、남경(3주)、복주(1주)4개지구;장53주산매균주여대장애희균EC600진행접합,획득28주접합자;PFGE분형결과현시,폐염극뢰백균(33주)가분위3개형2개아형、불로지구연산간균(9주)가분위4개형、음구장간균(5주)가분위4개형、대장애희균(6주)균위동일형;29주산KPC-2형매폐염극뢰백균적MLST분형서렬균위ST11,4주산IMP-4형매폐염극뢰백균중3주위ST876,1주위ST147.결론 재대아국11개성시적내약균감측중지유북경、항주、남경화복주적의원중검출휴대탄청매희매기인적균주,기중KPC-2최상견,기차위IMP-4、IMP-8,균주간동원성교고,유우탄청매희매기인재균주문용역발생수평전이,응인기림상고도관주.
Objective To analyze the genotype and molecular epidemiology of carbapenem-resistant Enterobacteriaceae.Method A total of 201 carbapenem-resistant Enterobacteriaceae were isolated from 14hospitals in 11 cities.The MICs of 14 antimicrobial drugs were detected using agar dilution method.Phenotypes of carbapenemases were screened using modified Hodge test and ethylene diamine tetraacetic acid (EDTA) test.Drug resistance genes were screened using PCR method.The strains carrying carbapenem resistance genes were confirmed by conjugation test.Homology analysis was carried out using pulsed-field gel electro-phoresis (PFGE) method and the epidemiological correlation is analyzed based on the Multilocus Sequence Typing (MLST) method in order to study the molecular epidemiology of carbapenem-resistant Enterobacteriaceae.Results Fifty-three strains among 201 carbapenem-insensitive Enterobacteriaceae were detected positive carbapenem-resistant genes,among which included 33 Klebsiella pneumoniae,9 Citrobacter freundii,6 Escherichia coli and 5 Enterobacter cloacae.Among the 53 strains,43 were from Beijing,6 strains from Hangzhou,3 strains from Nanjing and one from Fuzhou.Resistance genes-harboring plasmids were successfully transferred from 28 of 53 strains to Escherichia coli EC600.The PFGE spectmm showed that 33 Klebsiella pneumoniae were classified into three types,9 Citrobacterfreundii classified into four types,5 Enterobacter cloacae classified into four types,while 6 Escherichia coli were the same type.Based on the results of MLST test,29 Klebsiella pneumoniae strains producing KPC-2 type carbapenemase were all ST11,while among the four Klebsiella pneumonia carrying IMP-4 carbapenem resistant gene,three strains were ST876,one was ST147.Conclusions Carbapenem-resistant genes were detected only in hospitals from Beijing,Hangzhou,Nanjing and Fuzhou,and type KPC-2 was the most common,followed by IMP-4 and IMP-8.High homology of resistant strains could be related to horizontal transfer of carbapenemase genes,which should cause great concern.