[1]张雨菡,姚杰,陆承平,等.人工诱导猪链球菌氟喹诺酮耐药株的靶位突变分析[J].南京农业大学学报,2009,32(4):133-137.[doi:10.7685/j.issn.1000-2030.2009.04.025]
 ZHANG Yu-han,YAO Jie,LU Cheng-ping,et al.Mutation analysis of induced fluoroquinolone-resistant mutants of Streptococcus suis in vitro[J].Journal of Nanjing Agricultural University,2009,32(4):133-137.[doi:10.7685/j.issn.1000-2030.2009.04.025]
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人工诱导猪链球菌氟喹诺酮耐药株的靶位突变分析()
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《南京农业大学学报》[ISSN:1000-2030/CN:32-1148/S]

卷:
32卷
期数:
2009年4期
页码:
133-137
栏目:
OA栏目
出版日期:
2009-11-30

文章信息/Info

Title:
Mutation analysis of induced fluoroquinolone-resistant mutants of Streptococcus suis in vitro
作者:
张雨菡姚杰陆承平王丽平
南京农业大学动物医学院,江苏南京210095
Author(s):
ZHANG Yu-han YAO Jie LU Cheng-ping WANG Li-ping
College of Veterinary Medicine, Nanjing Agricultural University, Nanjing 210095, China
关键词:
恩诺沙星 环丙沙星 猪链球菌2 DNA回旋酶 拓扑异构酶Ⅳ 靶位突变
Keywords:
enrofloxacin ciprofloxacin Streptococcus suis type 2 DNA gyrase topoisomerase Ⅳ target mutation
分类号:
R378.12
DOI:
10.7685/j.issn.1000-2030.2009.04.025
文献标志码:
A
摘要:
以4株临床分离的对环丙沙星和恩诺沙星敏感的猪链球菌2型菌株为研究对象,采用体外递增药物浓度的方法分别诱导了其对环丙沙星和恩诺沙星耐药的菌株,按CLSI推荐方法测定了环丙沙星和恩诺沙星对亲本敏感株和诱导耐药株的MIC,测定了亲本株和诱导耐药株的生长曲线,并采用PCR和基因测序的方法分析了诱导耐药株的DNA回旋酶(GyrA和GyrB)和拓扑异构酶Ⅳ(ParC和ParE)耐药决定区(QRDR)的基因突变和氨基酸序列变化。结果表明:浓度递增法成功诱导了猪链球菌对环丙沙星和恩诺沙星耐药性,其MIC分别由0.5 mg.L-1上升至128 mg.L-1;与敏感株比较,恩诺沙星与环丙沙星诱导的耐药菌在gyrA和gyrB,或parC和parE耐药决定区的氨基酸序列有突变,除了已报道的与氟喹诺酮耐药相关的ParC的Ser79Phe,GyrA的Ser81Arg,GyrB的Asp315Asn、Ser285Leu和Glu354Lys及ParE的Pro278Ser点突变外,在诱导菌中还出现了一些不曾报道的突变位点和氨基酸缺失,如GyrA的Gln118His和ParE的Asn297Tyr突变,GyrB的288~291位和ParC的62位氨基酸缺失。结果提示:逐步增加药物浓度可以诱导猪链球菌对氟喹诺酮类抗菌药耐药性,并导致主要靶位发生突变。
Abstract:
To perform a systematic analysis of point mutations in the quinolone resistance determining regions(QRDRs)of the DNA gyrase and topoisomerase genes of Streptococcus suis type 2 strain after in vitro exposure to stepwise increasing concentrations of enrofloxacin and ciprofloxacin. Four parent strains of S. suis type 2 were chosen for stepwise exposure to increasing levels of enrofloxacin and ciprofloxacin followed by selection of resistant mutants. The QRDRs of gyrA, gyrB, parC and parE correlating to mutants with increased MICs were analysed for point mutations. Multiple mutants with significantly increased MICs(128 mg·L-1)were generated from each parent strain. Most of the amino acid substitutions identified were Ser79Phe of ParC, Ser81Arg of GyrA, Asp315Asn, Ser285Leu, Glu354Lys of GryB and Pro278Ser of ParE, which were consistent to the mechanisms of resistance reported in clinical isolates of S. pneumoniae. However, it has some mutable points Gln118His of GyrA, Asn297Tyr of ParE as well as the deletions 288-291 amino acids of GyrB and 62 amino acid of ParC, which haven′t been documented and whether these nuleotide mutations associated with fluoroquinolone resistance are not clear. The number of induction/selection cycles required for the emergence of key point mutations in gyrA and parC was variable among strains. The results indicate that stepwise increasing concentration of enrofloxacin and ciprofloxacin can easily select resistant strains of S. suis.

参考文献/References:

[1]Aarestrup,F,M,Artursson,K,Artursson,K. Trends in the resitance to antimicrobial agents of Streptococcus suis isolates from Denmark and Sweden[J]. Veterinary Microbiology, 1998
[2]Wimenburg,K,S,Zadoks,R,N,Zadoks,R,N. Human Streptococcus suis meningitis in the United States[J]. New England Journal of Medicine, 2006
[3]Drlica,K. DNA gyrase,topoisomerase Ⅳ,and the 4-quinolones[J]. Microbiol Mol Rev, 1997
[4]Balsalobre,L,Liares,J,Liares,J. Viridans group streptococci are donors in horizontal transfer of topoisomerase Ⅳ genes to Streptococcus pneumoniae[J]. Antimicrobial Agents and Chemotherapy, 2003
[5]Kawamura,Y,Mishima,N,Mishima,N. First Streptococcus agalactiae isolates highly resistant to quinolones with point mutations in gyrA and parC[J]. Antimicrobial Agents and Chemotherapy, 2003
[6]Orscheln,R,C,Olson,S,M,Olson,S,M. Intrinsic reduced susceptibility of serotype 6 Streptococcus pyogenes to fluoquinolone antibiotics[J]. Journal of Infectious Diseases, 2005
[7]Alonso,R,Courvalin,P,Courvalin,P. parC mutation conferring ciprofloxacin resistance in Streptococcus pyogenes BM4513[J]. Antimicrobial Agents and Chemotherapy, 2002
[8]MacGowan,A,P,Wootton,M,Wootton,M. Exploration of the in vitro pharmacodynamic activity of moxifloxacin for Staphylococcus aureus and streptococci of Lancefield Groups A and G[J]. Journal of Antimicrobial Chemotherapy, 1999
[9]Clinical,and,Laboratory,Standards,Institute. Performance Standards for Antimicrobial Disk and Dilution Susceptibility Tests for Bacteria Isolated from Animals;Approved Standard[M]. Wayne,USA:CLSI, 2008
[10]Perichon,B,Courvalin,P,Courvalin,P. Characterization of a mutation in parE gene that confers fluoroquinolone resistance in Streptococcus pneumoniae[J]. Antimicrobial Agents and Chemotherapy, 1997
[11]Munoz,B. ParC subunit of DNA topoisomerase IV of Streplocorcus pneumoniae is a primary target of fluoroquinolones and cooperates with DNA gyrase A subunit in forming resistance phenotype[J]. Antimicrobial Agents and Chemotherapy, 1996
[12]Pan,X,S,Mehtar,S,Mehtar,S. Involvement of topoisomerase Ⅳ and DNA gyrase as ciprofloxacin targets in Stretococcus pneumoniae[J]. Antimicrobial Agents and Chemotherapy, 1996
[13]Tankovic,J,Duval,J,Duval,J. Contribution of mutations in gyrA and parC genes to fluoroquinolone resistance of mutants of Streptococcus pneumoniae obtained in vivo and in vitro[J]. Antimicrobial Agents and Chemotherapy, 1996
[14]Gootz,T,D,Haskell,S,Haskell,S. Activity of the new fluoroquinolone trovafloxacin(CP-99,219)against DNA gyrase and topoisomerase Ⅳ mutants of Streptococcus pneumoniae selected in vitro[J]. Antimicrobial Agents and Chemotherapy, 1996
[15]Weigel,L,M,Facklamr,R,Facklamr,R. Genetic analyses of mutations contributing to fluoroquinolone resistance in clinical isolates of Streplococcus pneumoniae[J]. Antimicrobial Agents and Chemotherapy, 2001
[16]Zhanel,G,G,Nichol,K,Nichol,K. Molecular characterization of fluoroquinolone resistant Streptococcus pneumoniae clinical isolates obtained from across Canada[J]. Diagnostic Microbiology and Infectious Disease, 2003
[17]Jones,M,E,Martin,N,Martin,N. Prevalence of gyrA,gyrB,parC and parE mutations in clinical isolates of Streptococcus pneumoniae with decreased susceptibilities to different fluoroquinolones and originating from worldwide surveillance studies during the 1997-1998 respiratory season[J]. Antimicrobial Agents and Chemotherapy, 2000
[18]Bast,D,J,Duncan,C,L,Duncan,C,L. Fluoroquinolone resistance in clinical isolates of Streptococcus pneumoniae:contributions of type Ⅱ topoisomerase mutations and efflux to levels of resistance[J]. Antimicrobial Agents and Chemotherapy, 2000
[19]Janoir,C,Kitzis,M,D,Kitzis,M,D. New mutation in ParE in a Pneumococcal in vitro mutant resistant to fluoroquinolones[J]. Antimicrobial Agents and Chemotherapy, 2001

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备注/Memo

备注/Memo:
公益性行业(农业)科研专项(200803016)
更新日期/Last Update: 2009-11-30