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Rapid Antibiotic Susceptibility Testing by Deuterium Labeling of Bacterial Lipids in On-Target Microdroplet Cultures
ACS Environmental Au Pub Date : 2022-05-27 , DOI: 10.1021/jasms.2c00056
Evan A Larson 1 , Josiah J Rensner 1 , Kristina R Larsen 2 , Bryan Bellaire 2 , Young Jin Lee 1
Affiliation  

Antimicrobial resistance is a serious challenge facing human and veterinary health. Current methods of detecting resistance are limited in turn-around time or universal detection. In this work, a new antimicrobial susceptibility test is developed and validated, which utilizes deuterium labeling of membrane lipids to track the growth of bacterial cells. We hypothesize that deuterium uptake and subsequent labeling of lipids can be detected using matrix-assisted laser desorption/ionization mass spectrometry (MALDI-MS). Additionally, bacteria growth is performed on the MALDI target, minimizing sample preparation materials and time. When two Escherichia coli strains are grown in the presence of deuterium oxide, labeling can be detected in as little as 30 min to 2 h. The labeling efficiency, or the ratio of labeled to unlabeled lipid peaks, provides information about the growth rate of bacteria. This growth ratio can differentiate between resistant and susceptible strains of bacteria as a resistant strain will maintain ∼50% labeling efficiency between untreated and treated cultures. In comparison, a susceptible strain will see a decrease in fractional abundance of deuterium from ∼50% in the untreated to ∼10% in the treated. This approach is applied to measure the minimum inhibitory concentration (MIC) of the resistant and susceptible strains from on-target microdroplet culture in a range of antibiotic concentrations. The first antibiotic concentration with a significant decrease in fractional abundance of deuterium correlates well with a traditionally obtained MIC using broth dilution, indicating the clinical relevance of the results.

中文翻译:

通过氘标记对靶向微滴培养物中的细菌脂质进行快速抗生素敏感性测试

抗菌素耐药性是人类和兽医健康面临的严峻挑战。当前检测电阻的方法在周转时间或通用检测方面受到限制。在这项工作中,开发并验证了一种新的抗菌药物敏感性测试,该测试利用膜脂的氘标记来跟踪细菌细胞的生长。我们假设可以使用基质辅助激光解吸/电离质谱 (MALDI-MS) 检测氘吸收和随后的脂质标记。此外,在 MALDI 目标上进行细菌生长,最大限度地减少样品制备材料和时间。当两个大肠杆菌菌株在氧化氘存在下生长,标记可以在短短 30 分钟到 2 小时内检测到。标记效率或标记与未标记脂质峰的比率提供了有关细菌生长速率的信息。这种生长比率可以区分耐药菌株和敏感菌株,因为耐药菌株将在未处理和处理过的培养物之间保持约 50% 的标记效率。相比之下,易感菌株将看到氘的丰度分数从未处理的约 50% 下降到处理后的约 10%。该方法适用于在一系列抗生素浓度下测量来自靶向微滴培养的耐药和敏感菌株的最小抑制浓度 (MIC)。
更新日期:2022-05-27
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