New Bioluminescence‑Based Antimicrobial Susceptibility Test: ATP as a Core Biomarker for Rapid Susceptibility Assays

A proof‑of‑concept study published in Frontiers in Microbiology has developed a bioluminescence‑based rapid antimicrobial susceptibility testing (AST) approach centered on ATP consumption. Leveraging the metabolic features of ATP, this method greatly shortens assay time and addresses the long‑standing clinical bottleneck of time‑consuming conventional AST. Conventional phenotypic AST requires 16‑20 hours of incubation, which may delay targeted anti‑infective therapy for critically‑ill infected patients.

Different from prior assays measuring intracellular bacterial ATP, this technique uses exogenous ATP as a detection probe. Resistant bacteria survive and multiply, consuming exogenous ATP in the system and yielding faint luminescence. By contrast, antibiotic‑susceptible bacteria are growth‑inhibited, leaving ATP preserved in the system. Strong bioluminescence is triggered when luciferin‑luciferase complex is added, enabling differentiation between resistant and susceptible phenotypes. In tests on 56 clinical Enterobacterales isolates across 348 assay sets, it achieved an overall concordance rate of 83.3 % versus the EUCAST disk‑diffusion method, with major errors at only 1.7 %. The whole assay can be finished within three hours.

While the ATP‑based probe eliminates bacterial lysis steps, limitations remain. The luminescent signal from the ATP‑luciferin system decays rapidly. Variable ATP‑consumption rates across bacterial strains cause result discrepancies, and luciferase requires cold‑chain storage.

Industry analysts note this work validates the feasibility of exogenous ATP as an AST biomarker. Further work includes expanding strain panels, improving reagent stability, developing lyophilized pre‑cast plates, and extending assays to Mycobacterium tuberculosis and fungi, to translate the ATP bioluminescence assay into routine clinical microbiology testing.

FOR DETAIL:  https://doi.org/10.3389/fmicb.2024.1357680

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