Please wait a minute...
Journal of ZheJiang University(Medical Science)  2013, Vol. 42 Issue (2): 149-155    DOI: 10.3785/j.issn.1008-9292.2013.02.004
    
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
Download: HTML (   PDF(1002KB)
Export: BibTeX | EndNote (RIS)      

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 wordsEnterobacter/drug effects      Acinetobacter baumannii/drug effects      Escherichia/drug effects      Drug resistance,bacterial      Enteric bacilli      Drug resistance      Integron      Drug resistance gene cassette      Evolution     
Received: 28 December 2012      Published: 25 March 2013
CLC:  R 378.2  
Cite this article:

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.

URL:

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


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

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 

[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. 
[1] WANG Mengyan, ZHU Biao. Research progress on genes mutations related to sulfa drug resistance in Pneumocystis jirovecii[J]. Journal of ZheJiang University(Medical Science), 2017, 46(5): 563-569.
[2] GE Yumei, CHEN Xuebo, HUANG Yanying, LYU Huoyang, ZHAO Zhao, ZHOU Yonglie. Drug resistance and protoporphyrin ferrochelatase of Ralstonia mannitolilytica[J]. Journal of ZheJiang University(Medical Science), 2017, 46(4): 413-420.
[3] LI Yu, WANG Yueyue, WANG Haifeng, ZHANG Lingyu, DING Yongxing, CHEN Sulian, YANG Qingling, CHEN Changjie. Effects of lncRNA RP11-770J1.3 and TMEM25 expression on paclitaxel resistance in human breast cancer cells[J]. Journal of ZheJiang University(Medical Science), 2017, 46(4): 364-370.
[4] WANG Hebin,LI Yang,LIU Xingang,ZHOU Jun,WANG Qingqing,TANG Guping. Preparation, characterization and cytology study of Pluronic-PEI micelles[J]. Journal of ZheJiang University(Medical Science), 2017, 46(2): 127-133.
[5] LIN Haijiang, SHEN Weiwei, QIU Danhong, ZHOU Sujuan, WU Qionghai, DING Yingying, LIU Xing, HE Na. HIV related risky behavioral network and molecular epidemiology of HIV seroconcordant couples in Taizhou municipality of Zhejiang Province[J]. Journal of ZheJiang University(Medical Science), 2016, 45(3): 236-242.
[6] LIAO Wei-chao, HE Ying, WANG Bin-sheng, HUANG He. Everolimus combined with all-trans retinoid acid reverses drug resistance in acute promyelocytic leukemia NB4-R1 cells[J]. Journal of ZheJiang University(Medical Science), 2015, 44(5): 525-531.
[7] ZHAO Zun-lan, CAI Ying, WANG Yang-yang, XIA Chun-lei, LI Cong-xin, CHEN Su-lian, YANG Qing-ling, CHEN Chang-jie. Effects of miRNA-21 on paclitaxel-resistance in human breast cancer cells[J]. Journal of ZheJiang University(Medical Science), 2015, 44(4): 400-409.
[8] WU Yi-Fei, SUN Ai-Hua, ZHAO Jin-Fang, GE Yu-Mei, YAN Jie. Distribution of drug inactive enzyme genes in bacterial isolates and mechanism of its induction and inhibition[J]. Journal of ZheJiang University(Medical Science), 2013, 42(2): 131-140.
[9] . Research advances on mechanisms of chemoresistance in acute myeloid leukemia[J]. Journal of ZheJiang University(Medical Science), 2012, 41(5): 576-580.
[10] . Research progress on MKP-1 in tumor drug resistance[J]. Journal of ZheJiang University(Medical Science), 2012, 41(1): 111-116.
[11] . Anti-proliferation and chemo-sensitization effects of apigenin on human lung cancer cells[J]. Journal of ZheJiang University(Medical Science), 2011, 40(5): 508-514.
[12] . Down-regulation of Nrf2 enhances the cytotoxicity of Oxaliplatin to H460 cells[J]. Journal of ZheJiang University(Medical Science), 2010, 39(1): 11-16.
[13] Chen Jian, Yu Sufei. Detection of the E Coli and K Pneumoniae Strains' Resistance to the Third Generation Cephalosporins[J]. Journal of ZheJiang University(Medical Science), 1999, 28(3): 117-119.
[14] Huang Jianrong, Gan Mengjiu, Qian Wen, et al. NOSOCOMIAL INFECTION OF COAGULASE-NEGATIVE STAPHYLOCOCCI AND ITS DRUG RESISTANCE[J]. Journal of ZheJiang University(Medical Science), 1996, 25(3): 119-121.
[15] Yang Hua, Dong Haitao, Cao Jiang, et al. ANALYSIS OF MDR 1 GENE EXPRESSION BY REVERSE TRANSCRIPTION-POL YMERASE CHAIN REACTION[J]. Journal of ZheJiang University(Medical Science), 1995, 24(2): 49-51.