Phenotypic Profiling of Antibiotic Response Signatures in Escherichia coli Using Raman Spectroscopy
Phenotypic Profiling of Antibiotic Response Signatures in Escherichia coli Using Raman Spectroscopy
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DOI：
10.1128/aac.02098-13
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发表时间：
2014-03-01
影响因子：
4.9
通讯作者：
Senger, R. S.
中科院分区：
文献类型：
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作者：
Athamneh, A. I. M.;Alajlouni, R. A.;Senger, R. S.
确定新的潜在抗生素的作用机制是一个必要但耗时和昂贵的过程。表型图谱已经被有效地用来帮助发现未表征药物的作用机制和分子靶点。在这项研究中，拉曼光谱被用来描述大肠杆菌对应用抗生素的表型反应。拉曼光谱的使用是有利的，因为它是非侵入性的，无标记，易于自动化，并且其结果可以实时获得。在这项研究中，大肠杆菌培养物受到15种不同抗生素(代表5种功能抗生素类别)的3倍MIC作用30分钟，然后用拉曼光谱(使用532 nm激发波长)进行分析。由此产生的拉曼光谱包含了足够的生化信息，以区分属于同一类别的单个抗生素诱导的图谱。收集的光谱数据被用来建立判别分析模型，以识别未知抗生素化合物对大肠杆菌培养物表型的影响。化学计量学分析表明，拉曼光谱能够预测未知抗生素的功能类别，并识别引起类似表型反应的单个抗生素。这项研究的结果证明了拉曼光谱作为一种细胞表型分析方法的能力及其对抗生素药物开发研究的潜在影响。
Identifying the mechanism of action of new potential antibiotics is a necessary but time-consuming and costly process. Phenotypic profiling has been utilized effectively to facilitate the discovery of the mechanism of action and molecular targets of uncharacterized drugs. In this research, Raman spectroscopy was used to profile the phenotypic response of Escherichia coli to applied antibiotics. The use of Raman spectroscopy is advantageous because it is noninvasive, label free, and prone to automation, and its results can be obtained in real time. In this research, E. coli cultures were subjected to three times the MICs of 15 different antibiotics (representing five functional antibiotic classes) with known mechanisms of action for 30 min before being analyzed by Raman spectroscopy (using a 532-nm excitation wavelength). The resulting Raman spectra contained sufficient biochemical information to distinguish between profiles induced by individual antibiotics belonging to the same class. The collected spectral data were used to build a discriminant analysis model that identified the effects of unknown antibiotic compounds on the phenotype of E. coli cultures. Chemometric analysis showed the ability of Raman spectroscopy to predict the functional class of an unknown antibiotic and to identify individual antibiotics that elicit similar phenotypic responses. Results of this research demonstrate the power of Raman spectroscopy as a cellular phenotypic profiling methodology and its potential impact on antibiotic drug development research.