Rapid Detection of Carbapenemase-Producing Organisms Using a Luminescent Biosensor

ABSTRACT Carbapenemase-producing organisms (CPOs) pose an urgent global health threat due to their ability to inactivate carbapenems, a group of last resort antibiotics. Infections caused by these pathogens are associated with poor patient outcomes, high mortality rates, and added burden to infection prevention and control programs, making early detection vital to ensure optimal antimicrobial therapy and appropriate implementation of infection control practices. In this study, we report the application of a luminescent whole-cell biosensor for the rapid detection of CPOs. This biosensor provides positive test results within 2.5 hours, inclusive of setup time, and has been validated with a panel of laboratory and clinical isolates producing a diverse range of carbapenemases (KPC, NDM, IMP, VIM and OXA-48-like). The assay successfully identified all CPO isolates tested with a sensitivity of 100%, including strains producing weak OXA-48-like carbapenemases which are sometimes missed by currently used detection methods. The assay also demonstrated a specificity of 100 %, with all non-CPO clinical isolates testing negative under the optimized assay conditions. Due to the rapid time-to-positivity, minimal setup requirements, and high sensitivity, this test could serve as an attractive alternative to CPO detection methods currently employed by clinical microbiology labs, and could also facilitate CPO screening in other settings ( e.g., environmental, agricultural).