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浙江大学学报(医学版)  2013, Vol. 42 Issue (2): 149-155    DOI: 10.3785/j.issn.1008-9292.2013.02.004
专题报道     
肠道杆菌耐药性及其耐药相关Ⅰ类整合子可变区结构与进化
王欢1,包其郁2,孙爱华3,赵金方4,葛玉梅5,严杰5
1.浙江省人民医院检验科,浙江 杭州 310014;
2.温州医学院检验医学院和生命科学学院,浙江 温州 325035;
3.浙江医学高等专科学校,浙江 杭州 310053;
4.浙江中医药大学附属第一医院检验科,浙江 杭州 310006;
5.浙江大学医学院病原生物学系,浙江 杭州 310058
Multidrug resistance of enteric bacilli and its relation to structure and molecular evolution of variable region in resistance-related class-Ⅰ integron
WANG Huan1, BAO Qi-Yu2, SUN Ai-Hua3, ZHAO Jin-Fang4, GE Yu-Mei5, YAN Jie5
1.Department of Laboratory Medicine,The People′s Hospital of Zhejiang Province,Hangzhou 310014,China; 2.School of Laboratory Medical Science and Life Science,Wenzhou Medical College,Wenzhou 325035,China; 3.Zhejiang Medical College,Hangzhou 310053,China; 4.Department of Laboratory Medicine,The First Affiliated Hospital of Zhejiang Chinese Medical University,Hangzhou 310006,China; 5.Department of Medical Microbiology and Parasitology,Zhejiang University School of Medicine,Hangzhou 310058,China
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摘要:

目的:了解大肠埃希菌、阴沟肠杆菌和鲍曼不动杆菌临床菌株耐药性,及其Ⅰ类整合子分布、可变区耐药基因盒与分子进化关系。
方法:采用K-B法检测大肠埃希菌、阴沟肠杆菌和鲍曼不动杆菌对12种药物的敏感性。采用PCR及其产物测序检测上述菌株中Ⅰ类整合子携带率及可变区耐药基因盒。采用Clustal X和MEGA软件分析Ⅰ类整合子基因盒中耐药基因分子进化关系。
结果:鲍曼不动杆菌对临床常用青霉素类和头孢菌素类抗生素耐药率为54.2%~100%,大肠埃希菌和阴沟肠杆菌对菌素类抗生素耐药率也高达41.6%~62.5%。62.5%(15/24)大肠埃希菌、67.9%(19/28)阴沟肠杆菌、83.3%(20/24)鲍曼不动杆菌检出Ⅰ类整合子,81.5%(44/54)Ⅰ类整合子呈现为4种单条带谱型,其余为3种双条带谱型。Ⅰ类整合子可变区耐药基因盒中存在aac(6′)、sad(3″)、aad(2″)、cat(4′)和dfr(7、A13和15)耐药基因,可分别介导细菌对氨基糖苷类、氯霉素和磺胺类抗生素耐药。根据二氢叶酸还原酶基因序列差异,上述菌株携带的Ⅰ类整合子可分成4个分子进化组。
结论:上述肠道杆菌耐药情况严重并携带高含多种耐药基因盒的Ⅰ类整合子,Ⅰ类整合子有4条不同的进化途径。

关键词: 肠杆菌属/药物作用鲍氏不动杆菌/药物作用埃希氏菌属/药物作用抗药性细菌肠道杆菌耐药整合子耐药基因盒进化    
Abstract:
Objective: To investigate the drug resistance of enteric bacilli and its relation to the drug resistance gene cassette in the variable region and molecular evolution of class-Ⅰ integron.
Methods: K-B assay was applied to measure the drug resistance of E.coli,E.cloacae and A.baumannii isolated against twelve antibiotics.The class-Ⅰ integron and drug resistance gene cassettes in the variable region of the integron were detected by PCR and sequencing of amplification products.The molecular evolution of drug resistance genes in the class-Ⅰ integrons was analyzed using Clustal X and MEGA software.
Results: 54.2%-100% of A.baumannii isolates were resistant to the penicillin and cephem antibiotics,while E.coli and E.cloacae isolates had resistance rates of 41.6%-62.5% to cephem antibiotics.62.5%(15/24) of E.coli,67.9%(19/28) of E.cloacae and 83.3%(20/24) of A.baumannii isolates were positive for class-Ⅰ integrons.81.5%(44/54) of class-Ⅰ integrons showed 4 different single band spectrums and the other class-Ⅰ integrons displayed 3 different double band spectrums.In the drug resistance gene cassettes in variable regions of class-Ⅰ integrons there were 7 types in 4 groups of drug resistance genes,including aac(6′), sad(3″),aad(2″),cat(4′) and dfr(types 7,A13 and 15), which induced the resistance to aminoglycosides and sulfamido antibiotics and chloromycin.The class-Ⅰ integrons in the isolates might be divided into 4 molecular evolution groups according to the diversity of dihydrofolate reductase encoding gene sequences.
Conclusion: The enteric bacilli have a high drug resistance and frequently carry class-Ⅰ integrons with 7 drug resistance gene cassettes which present 4 different evolutionary pathways.
Key words: Enterobacter/drug effects    Acinetobacter baumannii/drug effects    Escherichia/drug effects    Drug resistance,bacterial    Enteric bacilli    Drug resistance    Integron    Drug resistance gene cassette    Evolution
收稿日期: 2012-12-28 出版日期: 2013-03-25
:  R 378.2  
基金资助:

浙江省自然科学基金资助项目(LY12H19002);浙江省医药卫生科技计划资助项目(2011KYA005).

通讯作者: 严杰(1956-),男,教授,博导,医学博士,主要从事病原菌致病与耐药机制及相关新型药物与疫苗研究     E-mail: med_bp@zju.edu.cn
作者简介: 王欢(1981-),医学博士,从事病原菌致病与耐药机制研究.
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引用本文:

王欢, 包其郁, 孙爱华, 赵金方, 葛玉梅, 严杰. 肠道杆菌耐药性及其耐药相关Ⅰ类整合子可变区结构与进化[J]. 浙江大学学报(医学版), 2013, 42(2): 149-155.

WANG Huan, BAO Qi-Yu, SUN Ai-Hua, ZHAO Jin-Fang, GE Yu-Mei, YAN Jie. Multidrug resistance of enteric bacilli and its relation to structure and molecular evolution of variable region in resistance-related class-Ⅰ integron. Journal of ZheJiang University(Medical Science), 2013, 42(2): 149-155.

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http://www.zjujournals.com/med/CN/10.3785/j.issn.1008-9292.2013.02.004        http://www.zjujournals.com/med/CN/Y2013/V42/I2/149


[1]LESTARI E S,SEVERIN J A,VERBRUGH H A.Antimicrobial resistance among pathogenic bacteria in Southeast Asia
[J].Southeast Asian J Trop Med Public Health,2012,43(2):385-422.

[2]SUN A H,FAN X L,GU Yi,et al.Predominant porB1A and porB1B genotypes and correlation of gene mutations with drug resistance in Neisseria gonorrhoeae isolates in Eastern China
[J].BMC Infect Dis,2010,10:323-331.

[3]CANTN R, COQUE T M, BAQUERO F.Multi-resistant Gram-negative bacilli: from epidemics to endemics
[J].Curr Opin Infect Dis, 2003,16(4):315-325.

[4]XIAO YONGhong,SHEN Ping,WEI Zeqing(肖永红,沈萍,魏泽庆,等),et al.Mohnarin report 2010: Surveillance of bacterial resistance in East China
[J].Chinese Journal of Nosocomiology(中华医院感染学杂志),2011,21(23):4896-4902.(in Chinese)

[5]VEEN E L,SCHILDER A G M,TIMMERS T K,et al.Effect of long-term trimethoprim / sulfameth-oxazole treatment on resistance and integron prevalence in the intestinal flora: a randomized,double-blind,placebo-controlled trial in children
[J].J Antimicrob Chemother,2009,63(5):1011-1016.

[6]PAIRANO G,AGERSO Y,AARESTRUP F M,et al.Occurrence of integrons and resistance genes among sulphonam resistant Shigella spp.from Brazil
[J].Antimicrob Agents Chemother,2005,55(3):301-305.

[7]VO A T,van DUIJKEREN E,FLUIT A C,et al.Antibiotic resistance,integrons and Salmonella genomic island l among non-typhoidal Salmonella serovars in the the Netherlands
[J].Int J Antimicrob Agents,2006,28(3):172-179.

[8]FLUIT A C,SCHMITZ F J.Resistance integrons and super-integrons
[J].Clin Microbiol Infect,2004,10(4):272-288.

[9]SABAT M,PRATS G.Structure and function of integrons
[J].Enferm Infect Microbiol Clin,2002,20(7):341-345.

[10]ROE M T,VEGA E,PILLAI S D.Antimicrobial resistance markers of class 1 and class 2 integron- bearing Escherichia coli from irrigation water and sediments
[J].Emerg Infect Dis,2003,9(7):822-826.

[11]TURTON J F,KAUFMANN M E,GLOVER J,et al.Detection and typing of integrons in epidemic strains of Acinetobacter baumannii found in the United Kingdom
[J].J Clin Microbiol,2005,43(7):3074-3082.

[12]ROWE-MAGNUS D A,MAZEL D.The role of integron in antibiotic resistance gene capture
[J].Int J Med Microbiol,2002,292(2):115-125.

[13]DAZ-MEJA J J,AMBILE-CUEVAS C F,ROSA I,et al.An analysis of the evolutionary relationships of integron integrases,with emphasis on the prevalence of class 1 integrons in Escherichia coli isolates from clinical and environmental origins.Microbiology,2008,154(Pt 1):94-102.

[14]SAMBROOK J,FRITSCH E F,MANIATIS T.Molecular Cloning: A Laboratory Manual
[M].2nd edition,New York: Cold Sping Harbor Labooratory Press,1989,1.21-1.52,2.60-2.80,7.3-7.35,9.14-9.22.

[15]MEZZATESTA M L,GONA F,STEFANI S.Enterobacter cloacae complex: clinical impact and emerging antibiotic resistance
[J].Future Microbiol,2012,7(7):887-902.

[16]BEREKET W,HEMALATHA K,GETENET B,et al.Update on bacterial nosocomial infections
[J].Eur Rev Med Pharmacol Sci,2012,16(8):1039-1044.

[17]JEANA S S,HSUEH P R.High burden of antimicrobial resistance in Asia
[J].Int J Antimicrob Agents,2012,37(4):291-295.

[18]ZHUO Chao,Su Danhong,NI Yuxing(卓超,苏丹虹,倪语星,等),et al.CHINNET 2009 surveillance of antimicrobial resistance in E.coli and Klebsiella spp in China
[J].Chinese Journal of Infection and Chemotherapy(中国感染与化疗杂志),2010,10(6):430-435.(in Chinese)

[19]NANDI S,MAUEER J J,HOFACRE C,et al.Gram-positive bacteria are a major reservoir of class 1 antibiotic resistance integrons in poultry litter.Proc Natl Acad Sci USA,2004,101(18):7118-7122.

[20]PLOY M C,DENIS F,COURVALIN P,et al.Molecular characterization of integrons in Acinetobacter baumannii: descrip tion of a hybrid class 2 integron
[J].Antimicrob Agents Chemother,2000,44(10):2684-2688.

[21]YU Y,JI S,CHEN Y,et al.Resistance of strains producing extended-spectrum beta-lactamases and genotype distribution in China
[J].J Infect,2007,54(1):53-57.

[22]DULUCQ S,STONGE G,GAGNé V,et al.DNA variants in the dihydrofolate reductase gene and outcome in childhood ALL
[J].Blood,2007,111(7):3692-3700. 
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