A simple label-free method reveals bacterial growth dynamics and antibiotic action in real-time.
A simple label-free method reveals bacterial growth dynamics and antibiotic action in real-time.
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DOI：
10.1038/s41598-022-22671-6
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发表时间：
2022-11-12
影响因子：
4.6
通讯作者：
Gillespie, Stephen H.
中科院分区：
文献类型：
--
作者：
Hammond, Robert J. H.;Falconer, Kerry;Powell, Thomas;Bowness, Ruth;Gillespie, Stephen H.
由于检测细菌生长的方法相对不敏感，因此了解细菌对环境压力的反应受到阻碍。这意味着抗生素耐药性和其他生理学方法的研究通常需要24小时或更长时间。我们开发并测试了一种散射光和检测系统（SLIC）来解决这一挑战，建立了检测限和小接种物生长的阳性检测时间。我们比较了在不同的高和低营养液体培养基中生长的细菌的光散射和两种密切相关的生物体的生长动力学。使用Gompertz和Broken Stick方程对散射数据进行建模。细菌也暴露于美罗培南，庆大霉素和头孢西丁在一定范围内的浓度和液体培养物的光散射被实时捕获。我们确定SLIC的检测限为1-2 mL体积中10 - 100 cfu mL−1。在不到4小时内获得了不同营养物对细菌影响的定量测量，并且可以非常快速地区分具有BlaKPC β内酰胺酶的肺炎克雷伯氏菌1705与菌株1706的生长动力学差异。在超MIC浓度下测试的每种抗生素的作用速度存在剂量依赖性差异。真实的时间显示了庆大霉素的致死作用、美罗培南的溶解作用和头孢西丁的缓慢杀菌作用。值得注意的是，对抗生素敏感的菌株可以在几秒钟内被识别出来。这项研究表明了提高细菌检测灵敏度的重要性。这导致更快速地评估敏感性和捕获关于细菌生长动力学的丰富数据的能力。在超MIC浓度下发生的快速杀灭率，这是一个重要的发现，需要纳入药代动力学和药效学模型。重要的是，细菌检测灵敏度的提高使敏感性结果在几分钟内就可报告临床结果成为可能，正如我们已经证明的那样。
Understanding the response of bacteria to environmental stress is hampered by the relative insensitivity of methods to detect growth. This means studies of antibiotic resistance and other physiological methods often take 24 h or longer. We developed and tested a scattered light and detection system (SLIC) to address this challenge, establishing the limit of detection, and time to positive detection of the growth of small inocula. We compared the light-scattering of bacteria grown in varying high and low nutrient liquid medium and the growth dynamics of two closely related organisms. Scattering data was modelled using Gompertz and Broken Stick equations. Bacteria were also exposed meropenem, gentamicin and cefoxitin at a range of concentrations and light scattering of the liquid culture was captured in real-time. We established the limit of detection for SLIC to be between 10 and 100 cfu mL−1 in a volume of 1–2 mL. Quantitative measurement of the different nutrient effects on bacteria were obtained in less than four hours and it was possible to distinguish differences in the growth dynamics of Klebsiella pneumoniae 1705 possessing the BlaKPC betalactamase vs. strain 1706 very rapidly. There was a dose dependent difference in the speed of action of each antibiotic tested at supra-MIC concentrations. The lethal effect of gentamicin and lytic effect of meropenem, and slow bactericidal effect of cefoxitin were demonstrated in real time. Significantly, strains that were sensitive to antibiotics could be identified in seconds. This research demonstrates the critical importance of improving the sensitivity of bacterial detection. This results in more rapid assessment of susceptibility and the ability to capture a wealth of data on the growth dynamics of bacteria. The rapid rate at which killing occurs at supra-MIC concentrations, an important finding that needs to be incorporated into pharmacokinetic and pharmacodynamic models. Importantly, enhanced sensitivity of bacterial detection opens the possibility of susceptibility results being reportable clinically in a few minutes, as we have demonstrated.
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影响因子：
10
作者：
Wang, Fenglin;Raval, Yash;Chen, Hongyu;Tzeng, Tzuen-Rong J.;DesJardins, John D.;Anker, Jeffrey N.
通讯作者：
Anker, Jeffrey N.
影响因子：
2
作者：
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通讯作者：
Valls, Joaquim
影响因子：
3.7
作者：
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通讯作者：
Chuang HS
影响因子：
8.8
作者：
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通讯作者：
Cheang, Mary
影响因子：
3
作者：
Pope, C. F.;Gillespie, S. H.;McHugh, T. D.
通讯作者：
McHugh, T. D.