Multiple Reaction Monitoring Profiling (MRM-Profiling) of Lipids To Distinguish Strain-Level Differences in Microbial Resistance in Escherichia coli
Multiple Reaction Monitoring Profiling (MRM-Profiling) of Lipids To Distinguish Strain-Level Differences in Microbial Resistance in Escherichia coli
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DOI：
10.1021/acs.analchem.9b02465
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发表时间：
2019-09-03
影响因子：
7.4
通讯作者：
Cooks, R. Graham
中科院分区：
文献类型：
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作者：
Xie, Zhuoer;Gonzalez, Edwin;Cooks, R. Graham
全球范围内抗生素耐药性的增加是由于农业中抗生素的过度使用和药物中的过度处方造成的。为了提供适当和及时的患者支持，需要更快地诊断抗菌药物耐药性。目前的细菌鉴定方法依赖于基因组学和蛋白质组学，并使用与已知菌株数据库的比较，但代谢物和脂质的诊断价值尚未得到显着探索。标准质谱/色谱方法涉及样品制备和分离过程中的多次稀释。为了增加获取的化学信息量和脂质分析速度，应用了多反应监测分析（MRM-分析）。 MRM 分析工作流程包括发现阶段和筛选阶段。发现阶段采用前体 (PREC) 离子和中性丢失 (NL) 扫描来筛选具有代表性的合并样本，以查找与特定脂质类别相关的官能团。第一阶段的信息以前体/产物离子对 (MRM) 形式组织，筛选阶段快速询问单个样品的这些 MRM。在本研究中，我们对分别用阿莫西林或阿莫西林/克拉维酸（一种 β-内酰胺和 β-内酰胺酶抑制剂）培养的四种不同大肠杆菌菌株的脂质提取物进行了 MRM 分析。 t 检验、方差分析和受试者工作特征 (ROC) 曲线用于确定每个 MRM 的显着性。应用主成分分析来区分在允许或不允许细菌耐药性发展的条件下培养的不同菌株。结果表明，MRM 分析可区分耐药和非耐药大肠杆菌菌株的脂质谱。
The worldwide increase in antimicrobial resistance is due to antibiotic overuse in agriculture and overprescription in medicine. For appropriate and timely patient support, faster diagnosis of antimicrobial resistance is required. Current methods for bacterial identification rely on genomics and proteomics and use comparisons with databases of known strains, but the diagnostic value of metabolites and lipids has not been explored significantly. Standard mass spectrometry/chromatography methods involve multiple dilutions during sample preparation and separation. To increase the amount of chemical information acquired and the speed of analysis of lipids, multiple reaction monitoring profiling (MRM-Profiling) has been applied. The MRM-Profiling workflow includes a discovery stage and a screening stage. The discovery stage employs precursor (PREC) ion and neutral loss (NL) scans to screen representative pooled samples for functional groups associated with particular lipid classes. The information from the first stage is organized in precursor/product ion pairs, or MRMs, and the screening stage rapidly interrogates individual samples for these MRMs. In this study, we performed MRM-Profiling of lipid extracts from four different strains of Escherichia coli cultured with amoxicillin or amoxicillin/clavulanate, a beta-lactam and beta-lactamase inhibitor, respectively. t tests, analysis of variance and receiver operating characteristic (ROC) curves were used to determine the significance of each MRM. Principal component analysis was applied to distinguish different strains cultured under conditions that allowed or disallowed development of bacterial resistance. The results demonstrate that MRM-Profiling distinguishes the lipid profiles of resistant and nonresistant E. coli strains.