OBJECTIVES:Quinolone antimicrobials are frequently misused due to self-medication and suboptimal dose administration, leading to the development of resistance as well as treatment failure. The present study aimed to characterise plasmid-mediated quinolone resistance (PMQR) determinants and their genetic selection in the presence of quinolone stress within members of the Enterobacteriaceae. METHODS:A total of 209 non-duplicate Enterobacteriaceae isolates were collected from hospital and community health centres over the period July 2013-June 2014. Molecular characterisation of phenotypically screened quinolone-resistant isolates was done by multiplex PCR. Plasmids bearing the qnr and aac(6')-Ib-cr genes were transformed into Escherichia coli DH5α and were selected on Muller-Hinton agar plates containing 0.25μg/mL and 0.5μg/mL ciprofloxacin, norfloxacin, ofloxacin, levofloxacin and moxifloxacin. Conjugation experiments were performed to determine whether the aac(6')-Ib-cr- and qnr-carrying plasmids were self-transferable. RESULTS:The transformation assay revealed that transformants carrying qnrA could be selected in media containing norfloxacin, ciprofloxacin and levofloxacin, whereas qnrB and aac(6')-Ib-cr were selected on media containing norfloxacin and ciprofloxacin. Transformed qnrD could be selected in media containing norfloxacin and ofloxacin, and qnrS was selected only in the presence of levofloxacin. CONCLUSIONS:The presence of qnr genes has been associated with an increase in quinolone minimum inhibitory concentrations (MICs) and therefore leads to treatment failure when quinolones are used as selective therapeutic drugs. Since PMQR determinants have a high prevalence, effective measures should be taken and surveillance should be performed in order to avoid treatment failures using this group of antimicrobials.

译文

目的:喹诺酮类抗生素由于自我用药和次优剂量给药而经常被滥用,导致耐药性的发展以及治疗失败。本研究旨在表征质粒介导的喹诺酮耐药性(PMQR)决定簇及其在肠杆菌科成员体内存在喹诺酮胁迫下的遗传选择。
方法:在2013年7月至2014年6月期间,从医院和社区卫生中心收集了209株非重复性肠杆菌科细菌。通过多重PCR对表型筛选的喹诺酮耐药菌株进行了分子表征。将带有qnr和aac(6')-Ib-cr基因的质粒转化到大肠杆菌DH5α中,并在含有0.25μg/ mL和0.5μg/ mL环丙沙星,诺氟沙星,氧氟沙星,左氧氟沙星和莫西沙星的Muller-Hinton琼脂平板上进行选择。进行缀合实验以确定携带aac(6')-Ib-cr-和qnr的质粒是否可自我转移。
结果:转化试验表明,在含有诺氟沙星,环丙沙星和左氧氟沙星的培养基中可以选择携带qnrA的转化子,而在含有诺氟沙星和环丙沙星的培养基上选择qnrB和aac(6'-Ib-cr)。可以在包含诺氟沙星和氧氟沙星的培养基中选择转化的qnrD,仅在存在左氧氟沙星的情况下选择qnrS。
结论:qnr基因的存在与喹诺酮最小抑制浓度(MICs)的增加有关,因此当喹诺酮类药物用作选择性治疗药物时导致治疗失败。由于PMQR决定因素的患病率很高,因此应采取有效措施并进行监视,以避免使用这组抗生素治疗失败。

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