Rapid Detection of Carbapenemase-Producing Enterobacterales Using a Luminescent Whole-Cell Biosensor

Carbapenemase-producing Enterobacterales (CPE) 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 CPE. This biosensor provides positive test results within 2.5 h, 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 identified 81/83 CPE isolates tested with a sensitivity of 97.5%, 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-CPE 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 CPE detection methods currently employed by clinical microbiology laboratories and could also facilitate CPE screening in other settings (<i>e.g.,</i> environmental and agricultural).