成都地区人型支原体对喹诺酮类药物的耐药机制研究  

Resistance mechanism of Mycoplasma hominis to quinolones in Chengdu area

作　　者：张红艳 夏云[1] 赵鹃 丁少川 缪晓燕 Zhang Hongyan , Xia Yun, Zhao Juan, Ding Shaoehuan, Miao Xiaoyan(Department of Laboratory Medicine, the First Affiliated Hospital of Chongqing Medical University, Chongqing 400016, Chin)

机构地区：[1]重庆医科大学附属第一医院医学检验科,400016 [2]四川省医学科学院四川省人民医院(东院)检验科

出　　处：《中华检验医学杂志》2018年第5期385-389,共5页Chinese Journal of Laboratory Medicine

摘　　要：目的 检测成都地区人型支原体对喹诺酮类药物的耐药性,探索拓扑异构酶基因gyrA、gyrB、parC和parE突变与耐药性产生的相关机制,提供本地区耐药支原体的流行病学资料.方法 采用16SrRNA基因测序技术对支原体进行鉴定,微量肉汤稀释法进行抗生素敏感试验.耐药基因的扩增采用PCR方法,序列比对采用DNAMAN软件和BLAST进行分析.结果 人型支原体对环丙沙星、左氧氟沙星、莫西沙星和加替沙星的耐药率分别为92.4%（61/66）、87.9%（58/66）、71.2%（47/66）和66.7%（44/66）.筛选出对喹诺酮类药物呈现不同药敏表型的人型支原体45株进行拓扑异构酶基因的扩增测序,其中对莫西沙星和加替沙星耐药的31株均发生了GyrA S153L的氨基酸变异,变异率68.9%（31/45）;对环丙沙星和左氧氟沙星耐药的41株均发生了ParC S91I的氨基酸变异,变异率91.1%（41/45）.另外发现2株高水平耐药株发生了ParE A463S新的氨基酸变异,GyrB无氨基酸变异发生.结论 人型支原体对喹诺酮类的耐药主要与gyrA和parC基因突变导致的氨基酸改变密切相关,不同的喹诺酮类药物对人型支原体的作用靶位不同且高水平耐药与多个基因的突变有关.Objective To detect the resistance of Mycoplasma hominis to quinolones in Chengdu area,explore resistance mechanism of topoisomerase gene gyrA, gyrB, parC and parE mutations associated with drug resistance and provide epidemiological data.Methods Mycoplasma hominis was identified by 16SrRNA gene sequencing technique and antibiotic susceptibility test was carried out by broth microdilution method.Resistance genes were amplified by PCR,whereas sequence alignment was analyzed by DNAMAN software and BLAST.Results Resistance rates of Mycoplasma hominis to ciprofloxacin, levofloxacin, moxifloxacin and gatifloxacin were 92.4%（61/66）,87.9%（58/66）,71.2%（47/66）and 66.7%（44/66）,respectively.Totally 45 strains with different susceptibility to quinolones were screened for amplification and sequencing of topoisomerase genes, of which, 31 strains resistant to moxifloxacin and gatifloxacin harbored GyrA S153L amino acid mutation, 68.9%（31/45）, 41 strains resistant to ciprofloxacin and levofloxacin harbored ParC S91I amino acid mutation, 91.1%（41/45）.In addition, a new amino acid substitution of ParE A463S was found in 2 high-level resistant strains.No amino acid change was found in GyrB.Conclusions Resistance of Mycoplasma hominis to quinolones is closely associated with amino acid changes caused by mutations in gyrA and parC genes.Different quinolones have different targeting roles and high level resistance is associated with multiple gene mutations.

关 键 词：人型支原体 喹诺酮类 抗药性 细菌 DNA促旋酶 DNA拓扑异构酶Ⅳ 突变 

分 类 号：R446.5[医药卫生—诊断学]