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Phenotypic Antibiotic Susceptibility Testing at the limit of one bacterial cell

Ahmad, I.; Johansson, L.; Larsson, J.; Zikrin, S.; Knagge, P.; Fange, D.; Elf, J.

2025-04-13 microbiology
10.1101/2025.04.13.648565 bioRxiv
Show abstract

The blood of a septic patient contains only a few bacteria per milliliter. Recently, various techniques have been developed for extracting these few bacteria from a blood sample. Independent of how these bacteria are separated from the blood cells, we want to learn from them how to treat the infection. Here, we describe how a phenotypic Antibiotic Susceptibility Test can be executed with a single bacterial cell by making averages over time instead of populations, if we account for the experimental noise and cell-to-cell variability. We use the method to make preliminary estimates for how long it takes to distinguish a single susceptible from a resistant cell with statistical confidence. We also exemplify how it is possible to sequentially test different antibiotics, or different concentrations of the same antibiotic, on one cell lineage until a susceptible phenotype is detected. The assay can be followed by single-cell species identification using FISH probes.

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