In vitro evolution provides insights into mechanisms of Mycoplasma genitalium resistance to moxifloxacin.

IF 3.6 2区 医学 Q1 INFECTIOUS DISEASES
Teck-Phui Chua, Jose L Huaman, Jennifer Danielewski, Catriona S Bradshaw, Dorothy A Machalek, Michael J McDonald, Suzanne M Garland, Gerald L Murray
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引用次数: 0

Abstract

Background: Current knowledge of Mycoplasma genitalium fluoroquinolone resistance is based on sequence analyses of clinical samples, an approach with limited scope. Many potential resistance mutations have been described but their impact on moxifloxacin efficacy is unclear.

Objective: To investigate the impact of individual mutations on fluoroquinolone resistance through selection of moxifloxacin-resistant mutants in vitro.

Methods: M. genitalium G37 was passaged sequentially in sub-inhibitory concentrations of moxifloxacin. MIC values were determined for moxifloxacin, sitafloxacin, levofloxacin and ciprofloxacin. Bacterial populations were profiled using amplicon sequencing.

Results: Across three independent experiments, four moxifloxacin mutants were isolated, with mutations encoding the following protein sequence variations: (i) GyrA D99Y/ParE E468K, (ii) ParC S83I/GyrA D99Y/ParE E468K, (iii) ParC D87V/GyrB P462S and (iv) GyrA M95I/ParE E468dup. Moxifloxacin MICs were elevated 16- to 32-fold for mutants with a single GyrA or ParC variation, and 128-fold for the dual GyrA/ParC mutant. Sitafloxacin MICs were elevated but remained lower than moxifloxacin MICs. Mutations did not have a substantial impact on in vitro growth dynamics. Population analysis showed that multiple mutations attained detectable population-wide frequencies, with evidence of clonal interference dynamics, with a minority becoming fixed in the population.

Conclusion: Mutations in multiple genes conferring fluoroquinolone resistance appeared with regularity in vitro. Findings of an additive effect for ParC/GyrA changes, and greater effectiveness of sitafloxacin against resistant bacteria compared with moxifloxacin, are both consistent with clinical data. Improved understanding of fluoroquinolone resistance will inform the development of diagnostic assays predicting fluoroquinolone susceptibility.

体外进化提供了生殖支原体对莫西沙星耐药机制的见解。
背景:目前对生殖支原体氟喹诺酮类药物耐药性的认识是基于临床样本的序列分析,这种方法的范围有限。许多潜在的耐药突变已被描述,但它们对莫西沙星疗效的影响尚不清楚。目的:通过体外筛选莫西沙星耐药突变体,探讨个体突变对氟喹诺酮类药物耐药性的影响。方法:在莫西沙星亚抑制浓度下依次传代生殖支原体G37。测定莫西沙星、西他沙星、左氧氟沙星和环丙沙星的MIC值。利用扩增子测序对细菌种群进行分析。结果:通过三个独立的实验,分离到四个莫西沙星突变体,突变体编码以下蛋白质序列变异:(i) GyrA D99Y/ParE E468K, (ii) ParC S83I/GyrA D99Y/ParE E468K, (iii) ParC D87V/GyrB P462S和(iv) GyrA M95I/ParE E468dup。单GyrA或ParC突变体的莫西沙星mic升高16- 32倍,双GyrA/ParC突变体的mic升高128倍。西他沙星mic升高,但仍低于莫西沙星mic。突变对体外生长动力学没有实质性影响。种群分析表明,多个突变达到了可检测的种群范围频率,有证据表明存在克隆干扰动力学,少数突变在种群中变得固定。结论:体外引起氟喹诺酮类药物耐药的多个基因突变具有一定的规律性。研究发现,与莫西沙星相比,西他沙星对耐药菌具有更大的疗效,并且对ParC/GyrA变化具有叠加效应,这与临床数据一致。提高对氟喹诺酮类药物耐药性的了解将有助于开发预测氟喹诺酮类药物敏感性的诊断检测方法。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
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来源期刊
CiteScore
9.20
自引率
5.80%
发文量
423
审稿时长
2-4 weeks
期刊介绍: The Journal publishes articles that further knowledge and advance the science and application of antimicrobial chemotherapy with antibiotics and antifungal, antiviral and antiprotozoal agents. The Journal publishes primarily in human medicine, and articles in veterinary medicine likely to have an impact on global health.
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