Antibiotic Susceptibility Testing of the Gram-Negative Bacteria Based on Flow Cytometry

摘要: Rapidly treating infections with adequate antibiotics is of major importance. This requires a fast and accurate determination the antibiotic susceptibility bacterial pathogens. The most frequently used methods are slow because they based on measurement growth inhibition. Faster methods, such as PCR-based detection determinants resistance, do not always provide relevant information susceptibility, particularly that which genetically based. Consequently, new changes in physiology caused by using flow cytometry fluorescent viability markers, being explored. In this study, we assessed whether Alexa Fluor® 633 Hydrazide (AFH), targets carbonyl groups, can be for testing. Carbonylation cellular macromolecules, increases antibiotic-treated cells, appropriate to assess purpose it irreversible. We tested clinical isolates Gram-negative bacteria, Escherichia coli Pseudomonas aeruginosa, from three classes: β-lactams, aminoglycosides fluoroquinolones. addition AFH, TO-PRO®-3, enters cells damaged membranes binds DNA, DiBAC4 (3), depolarized membranes. also monitored antibiotic-induced morphological alterations analyzing light scattering signals. Although all dyes signals allowed antibiotic-sensitive AFH proved suitable reliable susceptibility.