氨基糖苷类耐药的肠杆菌科细菌16S rRNA甲基化酶基因研究

doi:10.3969/j.issn.1673-8640.2014.05.022

检验医学 ›› 2014, Vol. 29 ›› Issue (5): 528-534.DOI: 10.3969/j.issn.1673-8640.2014.05.022

氨基糖苷类耐药的肠杆菌科细菌16S rRNA甲基化酶基因研究

吴琼¹, 韩立中², 孙景勇², 倪语星², 陈敏³

1.上海交通大学附属第六人民医院检验科, 上海 200233;
2.上海交通大学医学院附属瑞金医院微生物科, 上海 200025;
3.上海市疾病预防控制中心微生物实验室, 上海 200336

收稿日期:2013-07-15 出版日期:2014-05-30 发布日期:2014-05-27
作者简介:吴琼, 女, 1977年生, 博士, 主管技师, 主要从事细菌耐药机制研究。
基金资助:
国家自然科学基金资助项目(81101282)

Research on 16S rRNA methylase in aminoglycoside-resistant Enterobacteriaceae

WU Qiong¹, HAN Lizhong², SUN Jingyong², NI Yuxing², CHEN Min³

1. Department of Clinical Laboratory, Shanghai Sixth People′s Hospital, Shanghai Jiaotong University, Shanghai 200233, China;
2.Department of Microbiology, Ruijin Hospital, Shanghai Jiaotong University School of Medicine, Shanghai 200025, China;
3.Shanghai Municipal Center for Disease Control and Prevention, Shanghai 200336, China

Received:2013-07-15 Online:2014-05-30 Published:2014-05-27

摘要/Abstract

摘要： 目的探讨临床分离的对氨基糖苷类耐药的肠杆菌科细菌产16S rRNA甲基化酶状况, 分析其分子流行趋势及其耐药性形成和传播的机制。方法采用纸片扩散法筛选庆大霉素和/或阿米卡星耐药的肠杆菌科细菌;采用聚合酶链反应(PCR)扩增16S rRNA甲基化酶基因、氨基糖苷修饰酶基因、β-内酰胺酶基因;采用质粒接合试验验证16S rRNA甲基化酶的转移性;应用脉冲场凝胶电泳(PFGE)对16S rRNA甲基化酶基因阳性菌株进行分型。结果 201株对庆大霉素和/或阿米卡星耐药的肠杆菌科细菌中共检出38株16S rRNA甲基化酶阳性株(armA基因16株, rmtB基因22株)。其中30株可通过接合试验将耐药质粒转移至受体菌。bla_CTX-M-14、bla_TEM-1和 bla_SHV-12可连同armA或rmtB分别转移到11、20 和7个接合子中。肺炎克雷伯菌、大肠埃希菌和阴沟肠杆菌分别被PFGE分为4、21和1个型别。结论本研究分离的肠杆菌科细菌16S rRNA甲基化酶以armA和rmtB为主要流行型别, 且后者分离率较高。该甲基化酶可导致氨基糖苷类高水平耐药, 而且酶编码基因位于质粒上, 具有转移性, β-内酰胺酶基因和氟喹诺酮耐药决定因子可随之一同转移。

关键词: 16SrRNA甲基化酶, 肠杆菌科细菌, 氨基糖苷类药物

Abstract: Objective To investigate the molecular epidemiological characterization and the drug resistance and prevalence mechanism of 16S rRNA methylase in aminoglycoside-resistant Enterobacteriaceae isolated clinically. Methods Gentamicin-and or amikacin-resistant Enterobacteriaceae were screened by disc diffusion method. 16S rRNA methylase genes, aminoglycoside modification enzyme genes and beta-lactamase genes were amplified by polymerase chain reaction(PCR). The conjugal transfer of aminoglycoside-resistant determination was performed. Pulsed-field gel electrophoresis (PFGE) was carried out to analyze genotyping. Results A total of 16 armA gene and 22 rmtB gene 16S rRNA methylase positive isolates were identified. Plasmid conjugation experiments were successful with 30 armA- or rmtB-positive isolates. bla_CTX-M-14, bla_TEM-1 and bla_SHV-12 co-transferred with armA or rmtB were transferred to 11, 20 and 7 isolates. Klebsiella pneumoniae, Escherichia coli and Enterobacter cloacae were divided into 4, 21 and 1 PFGE patterns. Conclusions armA and rmtB are the main types of 16S rRNA methylase in Enterobacteriaceae isolated in our hospital, and rmtB is more prevalent than armA. This 16S rRNA methylase could mediate high-level resistance to aminoglycoside, and the encoding genes are located on plasmid that could transfer among different species. Beta-lactamase genes and fluoroquinolone resistance could be cotransferred with armA or rmtB gene.

Key words: 16S rRNA methylase, Enterobacteriaceae, Aminoglycoside drug

中图分类号:

R378.2

吴琼, 韩立中, 孙景勇, 倪语星, 陈敏. 氨基糖苷类耐药的肠杆菌科细菌16S rRNA甲基化酶基因研究[J]. 检验医学, 2014, 29(5): 528-534.

WU Qiong, HAN Lizhong, SUN Jingyong, NI Yuxing, CHEN Min. Research on 16S rRNA methylase in aminoglycoside-resistant Enterobacteriaceae[J]. , 2014, 29(5): 528-534.

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氨基糖苷类耐药的肠杆菌科细菌16S rRNA甲基化酶基因研究

Research on 16S rRNA methylase in aminoglycoside-resistant Enterobacteriaceae

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