Rapid Antibiotic Susceptibility Determination by Fluorescence Lifetime Tracking of Bacterial Metabolism.

Autor:	Rojas-Andrade MD; Department of Materials Science and Engineering, University of California, Irvine, Irvine, California 92697, United States., Perinbam K; Department of Physics and Astronomy University of California, Irvine, Irvine, California 92697, United States., Nguyen QT; Department of Physics and Astronomy University of California, Irvine, Irvine, California 92697, United States., Kim JS; Department of Molecular Biology and Biochemistry, University of California, Irvine, Irvine, California 92697, United States., Palomba F; Department of Biomedical Engineering University of California, Irvine, Irvine, California 92697, United States., Whiteson K; Department of Molecular Biology and Biochemistry, University of California, Irvine, Irvine, California 92697, United States., Digman MA; Department of Biomedical Engineering University of California, Irvine, Irvine, California 92697, United States., Siryaporn A; Department of Physics and Astronomy University of California, Irvine, Irvine, California 92697, United States.; Department of Molecular Biology and Biochemistry, University of California, Irvine, Irvine, California 92697, United States., Hochbaum AI; Department of Materials Science and Engineering, University of California, Irvine, Irvine, California 92697, United States.; Department of Molecular Biology and Biochemistry, University of California, Irvine, Irvine, California 92697, United States.; Department of Chemistry, University of California, Irvine, Irvine, California, 92697, United States.; Department of Chemical and Biomolecular Engineering, University of California, Irvine, Irvine, California 92697, United States.
Jazyk:	angličtina
Zdroj:	ACS infectious diseases [ACS Infect Dis] 2024 Dec 13; Vol. 10 (12), pp. 4057-4065. Date of Electronic Publication: 2024 Nov 21.
DOI:	10.1021/acsinfecdis.4c00491
Abstrakt:	To combat the rise of antibiotic-resistance in bacteria and the resulting effects on healthcare worldwide, new technologies are needed that can perform rapid antibiotic susceptibility testing (AST). Conventional clinical methods for AST rely on growth-based assays, which typically require long incubation times to obtain quantitative results, representing a major bottleneck in the determination of the optimal antibiotic regimen to treat patients. Here, we demonstrate a rapid AST method based on the metabolic activity measured by fluorescence lifetime imaging microscopy (FLIM). Using lab strains and clinical isolates of Escherichia coli with tetracycline-susceptible and resistant phenotypes as models, we demonstrate that changes in metabolic state associated with antibiotic susceptibility can be quantitatively tracked by FLIM. Our results show that the magnitude of metabolic perturbation resulting from antibiotic activity correlates with susceptibility evaluated by conventional metrics. Moreover, susceptible and resistant phenotypes can be differentiated in as short as 10 min after antibiotic exposure. This FLIM-AST (FAST) method can be applied to other antibiotics and provides insights into the nature of metabolic perturbations inside bacterial cells resulting from antibiotic exposure with single cell resolution.
Databáze:	MEDLINE
Externí odkaz: