A Novel Diagnostic Molecule for Quantifying Efflux Levels in Gram-Positive Bacteria

Funding Information: All authors contributed equally to this work. This project was partially funded by the A*Midex Foundation Interdisciplinarity Grant \u201CElisa.\u201D M.P. was supported by the CNRS\u2010Innovation \u201CPhenux\u201D Grant. M.A. and A.V.V. gratefully acknowledge the \u201COne Health Basic and Translational Research Actions addressing Unmet Needs on Emerging Infectious Diseases (INF\u2010ACT)\u201D, PNRR, mission 4, component 2, investment 1.3, project number foundation by the Italian Ministry of University and Research PE00000007 (University of Cagliari). M.A. and A.V.V. received financial support from the NIAID/NIH grant no. R01AI136799. Additional support came from Funda\u00E7\u00E3o para a Ci\u00EAncia e a Tecnologia (FCT, Portugal) through funds to GHTM \u2013 UID/04413/2020, LA\u2010REAL \u2013 LA/P/0117/2020, to project BIOSAFE (c, PTDC/CAL\u2010EST/30713/2017), and to the Project DREBI, Ref. 2022.07931.PTDC ( http://doi.org/10.54499/2022.07931.PTDC ). CF was supported by grant UI/BD/154472/2022 from FCT, Portugal. SSC was supported by FCT through CEECINST/00042/2021/CP1773/CT0009 ( https://doi.org/10.54499/CEECINST/00042/2021/CP1773/CT0009 ). The authors are grateful to Michael J. Rybak, Wayne State University, USA, and Dr. Lorena Tuchscherr, Jena University Hospital, Germany, for providing strains of mepA and norA mutants, respectively. Special thanks to Jean\u2010Michel Jault and C\u00E9dric Orelle from Universit\u00E9 Lyon, France, for supplying PatA/PatB and BmrA efflux mutants. The authors also appreciate the encouragement of SATT\u2010SUD and Idylle Labs, Paris, for their assistance in transforming this concept. The authors thank Dr. V\u00E9ronique Sinou (MCT, AMU, Marseille) for providing ESKAPE reference bacteria and SAK1758 bacteria. The authors thank Paolo Ruggerone (University of Cagliari) for valuable discussions. Most importantly, the authors acknowledge the tireless work of the IHTM team in Lisbon. The authors also extend our gratitude to Serhii Krykun (L'Institut national des sciences appliqu\u00E9es de Rouen) and Simon Pascal (University of Nantes) for their technical assistance. S. aureus Publisher Copyright: © 2025 The Author(s). Advanced Healthcare Materials published by Wiley-VCH GmbH.