PDF(Pubmed)
Abstract：
Antimicrobial susceptibility tests (ASTs) are pivotal in combating multidrug resistant pathogens, yet they can be time-consuming, labor-intensive, and unstable. Using the AST of tigecycline for sepsis as the main model, here we establish an automated system of Clinical Antimicrobials Susceptibility Test Ramanometry (CAST-R), based on D2O-probed Raman microspectroscopy. Featuring a liquid robot for sample pretreatment and a machine learning-based control scheme for data acquisition and quality control, the 3-h, automated CAST-R process accelerates AST by >10-fold, processes 96 paralleled antibiotic-exposure reactions, and produces high-quality Raman spectra. The Expedited Minimal Inhibitory Concentration via Metabolic Activity is proposed as a quantitative and broadly applicable parameter for metabolism-based AST, which shows 99% essential agreement and 93% categorical agreement with the broth microdilution method (BMD) when tested on 100 Acinetobacter baumannii isolates. Further tests on 26 clinically positive blood samples for eight antimicrobials, including tigecycline, meropenem, ceftazidime, ampicillin/sulbactam, oxacillin, clindamycin, vancomycin, and levofloxacin reveal 93% categorical agreement with BMD-based results. The automation, speed, reliability, and general applicability of CAST-R suggest its potential utility for guiding the clinical administration of antimicrobials.
摘要：
抗菌药敏试验(ASTs)在对抗多重耐药病原体方面至关重要，然而它们可能很耗时，劳动密集型,和不稳定。以替加环素治疗脓毒症的AST为主要模型,在这里，我们建立了一个自动化的临床抗菌药物敏感性测试拉曼测定系统（CAST-R），基于D2O探测的拉曼光谱。具有用于样品预处理的液体机器人和用于数据采集和质量控制的基于机器学习的控制方案，3-h，自动化CAST-R工艺将AST加速>10倍，过程96平行的抗生素暴露反应，并产生高质量的拉曼光谱。通过代谢活性的加速最小抑制浓度被提出作为基于代谢的AST的定量和广泛适用的参数，在100个鲍曼不动杆菌分离株上进行测试时，与肉汤微量稀释法（BMD）显示出99％的基本一致性和93％的分类一致性。对26份临床阳性血液样本进行8种抗菌药物的进一步检测，包括替加环素,美罗培南,头孢他啶,氨苄西林/舒巴坦,苯唑西林,克林霉素,万古霉素,左氧氟沙星与基于BMD的结果具有93％的分类一致性。自动化,速度,可靠性,CAST-R的普遍适用性表明其在指导抗菌药物临床使用方面的潜在效用。
