中华微生物学和免疫学杂志
中華微生物學和免疫學雜誌
중화미생물학화면역학잡지
CHINESE JOURNAL OF MICROBIOLOGY AND IMMUNOLOGY
2012年
10期
861-865
,共5页
刘洋%李方去%蒋伟燕%杨锦红%李向阳
劉洋%李方去%蔣偉燕%楊錦紅%李嚮暘
류양%리방거%장위연%양금홍%리향양
肺炎克雷伯菌%分子流行病学%β-内酰胺酶类%耐药性
肺炎剋雷伯菌%分子流行病學%β-內酰胺酶類%耐藥性
폐염극뢰백균%분자류행병학%β-내선알매류%내약성
Klebsiella pneumoniae%Molecular epidemiology%β-lactamas%Drug resistance
目的 研究碳青霉烯类耐药肺炎克雷伯菌临床儿童分离株的耐药特点及分子流行病学特征.方法 收集温州医学院附属第二医院2010年7月-2011年6月从儿童标本中分离的耐碳青霉烯类肺炎克雷伯菌12株,所有菌株为非重复菌株,菌种鉴定采用全自动微生物分析仪.改良的Hodge试验筛选产碳青霉烯酶阳性菌株,采用PCR法检测KPC、IMP、bla(s)、VIM、SPM和整合酶基因,测序确定基因型.对菌株进行质粒结合试验、质粒消除试验检测质粒的转移性.脉冲场凝胶电泳(PFGE)分析耐药菌株的同源性.结果 12株耐碳青霉烯类肺炎克雷伯菌对庆大霉素、妥布霉素、阿米卡星、环丙沙星、左氧氟沙星、复方磺胺甲噁唑的敏感率分别为8.3％、41.7％、58.3％、8.3％、8.3％、33.3％；12株菌均携带有KPC-2基因,且同时携带有TEM-1和SHV型β-内酰胺酶基因,其中SHV-11-like和SHV-1 2-like各6株；11株携带CTX-M型基因,其中4株为CTX-M-14-like基因,6株CTX-M-15-like基因；2株携带有OXA-10型基因,1株携带有PER-1基因.未检出NDM-1、GIM、SPM、SIM、VIM型碳青霉烯酶基因.12株均为Ⅰ类整合酶基因(int1)阳性.2株通过接合试验把质粒传递给受体菌EC600.所有接合子blaTEM-1基因阳性、超广谱β-内酰胺酶(ESBL)基因阳性及对亚胺培南、庆大霉素、阿米卡星、妥布霉素和头孢噻肟耐药,接合子ESBL基因型与供菌一致.2株菌经质粒消除后对亚胺培南的MIC值均有较大程度降低,消除后KPC-2基因扩增为阴性.12株KPC-2基因阳性菌株经PFGE分成5个基因型,主要为B型和C型.结论 KPC-2型碳青霉烯酶基因已经在儿童肺炎克雷伯菌中播散,常伴随携带多种类型的ESBL基因和Ⅰ类整合酶基因,部分耐药基因可通过质粒播散.
目的 研究碳青黴烯類耐藥肺炎剋雷伯菌臨床兒童分離株的耐藥特點及分子流行病學特徵.方法 收集溫州醫學院附屬第二醫院2010年7月-2011年6月從兒童標本中分離的耐碳青黴烯類肺炎剋雷伯菌12株,所有菌株為非重複菌株,菌種鑒定採用全自動微生物分析儀.改良的Hodge試驗篩選產碳青黴烯酶暘性菌株,採用PCR法檢測KPC、IMP、bla(s)、VIM、SPM和整閤酶基因,測序確定基因型.對菌株進行質粒結閤試驗、質粒消除試驗檢測質粒的轉移性.脈遲場凝膠電泳(PFGE)分析耐藥菌株的同源性.結果 12株耐碳青黴烯類肺炎剋雷伯菌對慶大黴素、妥佈黴素、阿米卡星、環丙沙星、左氧氟沙星、複方磺胺甲噁唑的敏感率分彆為8.3％、41.7％、58.3％、8.3％、8.3％、33.3％；12株菌均攜帶有KPC-2基因,且同時攜帶有TEM-1和SHV型β-內酰胺酶基因,其中SHV-11-like和SHV-1 2-like各6株；11株攜帶CTX-M型基因,其中4株為CTX-M-14-like基因,6株CTX-M-15-like基因；2株攜帶有OXA-10型基因,1株攜帶有PER-1基因.未檢齣NDM-1、GIM、SPM、SIM、VIM型碳青黴烯酶基因.12株均為Ⅰ類整閤酶基因(int1)暘性.2株通過接閤試驗把質粒傳遞給受體菌EC600.所有接閤子blaTEM-1基因暘性、超廣譜β-內酰胺酶(ESBL)基因暘性及對亞胺培南、慶大黴素、阿米卡星、妥佈黴素和頭孢噻肟耐藥,接閤子ESBL基因型與供菌一緻.2株菌經質粒消除後對亞胺培南的MIC值均有較大程度降低,消除後KPC-2基因擴增為陰性.12株KPC-2基因暘性菌株經PFGE分成5箇基因型,主要為B型和C型.結論 KPC-2型碳青黴烯酶基因已經在兒童肺炎剋雷伯菌中播散,常伴隨攜帶多種類型的ESBL基因和Ⅰ類整閤酶基因,部分耐藥基因可通過質粒播散.
목적 연구탄청매희류내약폐염극뢰백균림상인동분리주적내약특점급분자류행병학특정.방법 수집온주의학원부속제이의원2010년7월-2011년6월종인동표본중분리적내탄청매희류폐염극뢰백균12주,소유균주위비중복균주,균충감정채용전자동미생물분석의.개량적Hodge시험사선산탄청매희매양성균주,채용PCR법검측KPC、IMP、bla(s)、VIM、SPM화정합매기인,측서학정기인형.대균주진행질립결합시험、질립소제시험검측질립적전이성.맥충장응효전영(PFGE)분석내약균주적동원성.결과 12주내탄청매희류폐염극뢰백균대경대매소、타포매소、아미잡성、배병사성、좌양불사성、복방광알갑오서적민감솔분별위8.3％、41.7％、58.3％、8.3％、8.3％、33.3％；12주균균휴대유KPC-2기인,차동시휴대유TEM-1화SHV형β-내선알매기인,기중SHV-11-like화SHV-1 2-like각6주；11주휴대CTX-M형기인,기중4주위CTX-M-14-like기인,6주CTX-M-15-like기인；2주휴대유OXA-10형기인,1주휴대유PER-1기인.미검출NDM-1、GIM、SPM、SIM、VIM형탄청매희매기인.12주균위Ⅰ류정합매기인(int1)양성.2주통과접합시험파질립전체급수체균EC600.소유접합자blaTEM-1기인양성、초엄보β-내선알매(ESBL)기인양성급대아알배남、경대매소、아미잡성、타포매소화두포새우내약,접합자ESBL기인형여공균일치.2주균경질립소제후대아알배남적MIC치균유교대정도강저,소제후KPC-2기인확증위음성.12주KPC-2기인양성균주경PFGE분성5개기인형,주요위B형화C형.결론 KPC-2형탄청매희매기인이경재인동폐염극뢰백균중파산,상반수휴대다충류형적ESBL기인화Ⅰ류정합매기인,부분내약기인가통과질립파산.
Objective To investigate molecular epidemiology and antimicrobial susceptibility of carbapenem-resistant strains of Klebsiella pneumoniae isolated from children.Methods From July 2010 to June 2011,twelve non-replicate clinical isolates of carbapenem-resistant Klebsiella pneumoniae were consecutively collected from children inpatients in the Second Hospital of Wenzhou Medical Colloge.All of the isolates were identified by the automated microbiology systems.Modified Hodge test was used to screen strains producing carbapenemases.Pulsed field gel electrophoresis(PFGE) was performed to analyze the homogeneity of genomic DNA of Klebsiella pneumoniae.KPC,IMP,GIM,SPM,SME,OXA-10,bla(s),VIM gene and integrase gene were amplified by PCR and then sequenced to cofirm the genotypes;Plasmid conjugation experiment was used to study the transfer method of bacterial resistance.Plasmid-curing test were used to initally locate the resistant genes.Results One(8.3％),5(41.7％),7(58.3％),1(8.3％),1(8.3％) and4(33.3％) of12isolates were susceptible to gentamicin,tobramycin,amikacin,ciprofloxacin,levofloxacin and trimoxazole,respectively.All isolates carried KPC-2,TEM-1 and SHV genes(six for SHV-11-like,six for SHV-12-like).Eleven of twelve isolates with KPC-2 gene carried CTX-M genes(4 for CTX-M-14-like,6 for CTX-M-15-like).Two isolates carried OXA-10 genes,and one isolates carried PER-1 gene.None of NDM-1,GIM,SPM,SIM and VIM carbapenemase genes was detected in 12 isolates.All of 12 isolates carried Int 1 genes.The plasmids of 2 isolates were transgerred into the recipients E.coli EC600.PCR and sequence analysis revealed that blaTEM-1 and blaCTX-M-15-like were co-transferred with the KPC-2 gene to the recipients.Elimination of KPC-2-encoding plasmid from Kp7 and Kp12 resulted in imipenem susceptibility in the two isolates.Amplification revealed that KPC-2 gene was lost by the plasmid-curing test.Of the 12 isolates,5 patterns were obtained by PFGE.Pattern B and C were the main drug resistant clones.Conclusion KPC-2 gene are the major carbapenemase genes in Klebsiella pneumoniae isolated from children,including ESBLs and integrase.Some resistance genes can be disseminated by plasmids.
