Liu Yi, Moore John H, Kolling Glynis L, McGrath John S, Papin Jason A, Swami Nathan S
Electrical and Computer Engineering, University of Virginia, Charlottesville, Virginia 22904, USA.
Biomedical Engineering, University of Virginia, Charlottesville, VA 22904, USA.
Sens Actuators B Chem. 2020 Jun 1;312. doi: 10.1016/j.snb.2020.127936. Epub 2020 Mar 4.
Infections due to () often exhibit broad-spectrum resistance and persistence to common antibiotics. Persistence is especially problematic with immune-compromised subjects who are unable to eliminate the inhibited bacteria. Hence, antibiotics must be used at the appropriate minimum bactericidal concentration (MBC) rather than at minimum inhibitory concentration (MIC) levels. However, MBC determination by conventional methods requires a 24 h culture step in the antibiotic media to confirm inhibition, followed by a 24 h sub-culture step in antibiotic-free media to confirm the lack of bacterial growth. We show that electrochemical detection of pyocyanin (PYO), which is a redox-active bacterial metabolite secreted by , can be used to rapidly assess the critical ciprofloxacin level required for bactericidal deactivation of within just 2 hours in antibiotic-treated growth media. The detection sensitivity for PYO can be enhanced by using nanoporous gold that is modified with a self-assembled monolayer to lower interference from oxygen reduction, while maintaining a low charge transfer resistance level and preventing electrode fouling within biological sample matrices. In this manner, bactericidal efficacy of ciprofloxacin towards at the MBC level and bacterial persistence at the MIC level can be determined rapidly, as validated at later timepoints using bacterial subculture in antibiotic-free media.
由()引起的感染通常对常见抗生素表现出广谱耐药性和持续性。对于无法清除受抑制细菌的免疫功能低下的受试者来说,持续性尤其成问题。因此,抗生素必须以适当的最低杀菌浓度(MBC)而非最低抑菌浓度(MIC)水平使用。然而,通过传统方法测定MBC需要在抗生素培养基中进行24小时培养步骤以确认抑制作用,随后在无抗生素培养基中进行24小时传代培养步骤以确认细菌生长的缺乏。我们表明,对绿脓菌素(PYO)进行电化学检测,绿脓菌素是由()分泌的一种具有氧化还原活性的细菌代谢产物,可用于在抗生素处理的生长培养基中仅2小时内快速评估杀菌失活所需的关键环丙沙星水平。通过使用用自组装单分子层修饰的纳米多孔金可以提高对PYO的检测灵敏度,以降低氧还原的干扰,同时保持低电荷转移电阻水平并防止生物样品基质中的电极污染。通过这种方式,可以快速确定环丙沙星在MBC水平对()的杀菌效力以及在MIC水平的细菌持续性,如在稍后时间点使用无抗生素培养基中的细菌传代培养所验证的那样。