Deuterium uptake in combination with Raman spectroscopy as a tool to investigate antibiotic susceptibility of bacteria
Deuterium uptake in combination with Raman spectroscopy as a tool to investigate antibiotic susceptibility of bacteria
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DOI：
10.1117/12.2553093
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发表时间：
2020-02
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通讯作者：
D. Bauer;L. Qiu;K. Wieland;Anna-Cathrine Neumann-Cip;A. Wieser;G. Magistro;C. Stief;C. Haisch
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作者：
D. Bauer;L. Qiu;K. Wieland;Anna-Cathrine Neumann-Cip;A. Wieser;G. Magistro;C. Stief;C. Haisch
多重耐药细菌的进化对我们全球的医疗体系构成了重大威胁。为了解决这一严重的公共卫生问题，需要显著提高最先进的抗生素敏感性测试(AST)的速度和灵敏度，如肉汤稀释法和纸片扩散法，以提供更好的患者护理。为了开发一种性能优于当前最先进程序的快速可靠的替代方案，开发了一种高通量、通常适用于除虫药物组合的分析方案，使用拉曼光谱作为非破坏性分析工具，以监测代谢活性细菌的吸氢情况，并另外提取细菌和/或污染物识别的特定化学信息。我们的AST是基于代表革兰氏阴性菌(大肠杆菌)和革兰氏阳性菌(粪肠球菌)的两个参考菌株，总共使用四种不同的抗生素治疗。除了高灵敏度和高特异度外，时间是临床诊断中最关键的参数。因此，对高度浓缩的样品进行批量分析比单电池方法更受青睐，以便能够及时、直接地进行样品制备和调查。开发的AST还包括预孵化步骤(在添加含有D2O的介质之前进行细菌药物孵化)，这被证明是开发由革兰氏阳性和革兰氏阴性细菌组成的可靠测试的关键。52株典型的尿路感染引起病原体的临床分离株在半自动环境中进行了研究，显示出与最先进的分析方法很好的一致性。
The evolution of multi-resistant bacteria poses a major threat to our healthcare system worldwide. To address this serious public health issue, the speed and sensitivity of state-of-the-art antibiotic susceptibility testing (AST) such as broth dilution and disc diffusion methods need to be significantly improved to provide better patient care. With the ultimate goal of developing a fast and reliable alternative that outperforms current state-of-the-art procedures, a high-throughput, generally– in terms of bug drug combinations – applicable analytical protocol was developed employing Raman spectroscopy as non-destructive analysis tool to monitor the deuterium uptake of metabolically active bacteria and additionally extract chemical-specific information for bacteria and/or contaminant identification. Our AST is based on two reference strains representing Gram-negative (E. coli) and Gram-positive bacteria (E. faecalis), treated with a total of four different antibiotics. Apart from high sensitivity and specificity, time is the most crucial parameter in clinical diagnosis. Hence, bulk analysis of highly concentrated samples was favored over a single-cell approach to allow timeefficient, straight-forward sample preparation and investigation. The developed AST also comprises a preincubation step (bug-drug incubation prior to the addition of D2O containing medium) which is shown to be key for the development of a reliable test comprising Gram-positive and Gram-negative bacteria. 52 clinical isolates typical for urinary tract infection causing pathogens were investigated in a semi-automated setting showing good agreement with state-of-the-art analytics.