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Using the AST of tigecycline for sepsis as the main model, here we establish an automated system of Clinical Antimicrobials Susceptibility Test Ramanometry (CAST\u2010R), based on D<jats:sub>2<\/jats:sub>O\u2010probed Raman microspectroscopy. Featuring a liquid robot for sample pretreatment and a machine learning\u2010based control scheme for data acquisition and quality control, the 3\u2010h, automated CAST\u2010R process accelerates AST by &gt;10\u2010fold, processes 96 paralleled antibiotic\u2010exposure reactions, and produces high\u2010quality Raman spectra. The Expedited Minimal Inhibitory Concentration via Metabolic Activity is proposed as a quantitative and broadly applicable parameter for metabolism\u2010based AST, which shows 99% essential agreement and 93% categorical agreement with the broth microdilution method (BMD) when tested on 100 <jats:italic>Acinetobacter baumannii<\/jats:italic> 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\u2010based results. The automation, speed, reliability, and general applicability of CAST\u2010R suggest its potential utility for guiding the clinical administration of antimicrobials.<\/jats:p>","DOI":"10.1002\/mlf2.12019","type":"journal-article","created":{"date-parts":[[2022,4,18]],"date-time":"2022-04-18T17:20:09Z","timestamp":1650302409000},"page":"329-340","update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":14,"title":["Rapid, automated, and reliable antimicrobial susceptibility test from positive blood culture by CAST\u2010R"],"prefix":"10.1002","volume":"1","author":[{"given":"Pengfei","family":"Zhu","sequence":"first","affiliation":[{"name":"Single\u2010Cell Center, CAS Key Laboratory of Biofuels, Shandong Key Laboratory of Energy Genetics and Shandong Energy Institute, Qingdao Institute of Bioenergy and Bioprocess Technology Chinese Academy of Sciences  Qingdao China"},{"name":"University of Chinese Academy of Sciences Beijing 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