Protocol to quantify bacterial burden in time-kill assays using colony-forming units and most probable number readouts for Mycobacterium tuberculosis
Here, we present a protocol to perform a time-kill assay (TKA) to quantify bacterial burden at multiple time points using colony-forming units and most probable number readouts simultaneously. We describe steps for preparing inoculum, experimental conditions, and sampling bacterial counts. We then detail procedures for quantification and analysis. TKAs provide longitudinal data reflecting the dynamics of the antibiotic effect over time against a planktonic culture and quantify the concentration-effect relationship. For complete details on the use and execution of this protocol, please refer to Van Wijk et al.<sup>1</sup>.
Authors
- Diana Angélica Aguilar-Ayala
- Jordana Galizia
- Laura Cioetto-Mazzabò
- Ainhoa Lucía (ORCID: https://orcid.org/0000-0002-0111-4697)
- Riccardo Manganelli (ORCID: https://orcid.org/0000-0003-0940-7336)
- Marie Sylvianne Rabodoarivelo (ORCID: https://orcid.org/0000-0002-4630-5446)
- Jan Rybniker (ORCID: https://orcid.org/0000-0001-8351-2690)
- Maria Rosalia Pasca (ORCID: https://orcid.org/0000-0002-8906-4937)
- Deborah Recchia (ORCID: https://orcid.org/0000-0002-2735-5165)
- Michael Dal Molin (ORCID: https://orcid.org/0000-0002-1587-6692)
- Lindsay Sonnenkalb (ORCID: https://orcid.org/0000-0002-8827-772X)
- Giulia Degiacomi (ORCID: https://orcid.org/0000-0002-3626-8311)
- Eik Hoffmann (ORCID: https://orcid.org/0000-0003-4224-1091)
- Santiago Ramón‐García (ORCID: https://orcid.org/0000-0002-8480-0325)
- Cyril Gaudin
Institutions
- University of Padua (IT)
- University of Cologne (DE)
- University of Pavia (IT)
- Universidad de Zaragoza (ES)
- Institut Pasteur de Lille (FR)
- Instituto de Salud Carlos III (ES)
- Fundacion Agencia Aragonesa para la Investigacion y el Desarrollo (ES)
- Center for Infection and Immunity of Lille (FR)
- Research Center Borstel - Leibniz Lung Center (DE)
Publication Details
- Journal
- STAR Protocols
- Published
- 2025-03-01
- DOI
- https://doi.org/10.1016/j.xpro.2025.103643
- Citations
- 5
- Primary Topic
- Tuberculosis Research and Epidemiology
- Type
- article
- Field-Weighted Citation Impact
- 6.98
Funders
- Bill and Melinda Gates Foundation
- University of Dundee
- European Federation of Pharmaceutical Industries and Associations
- Innovative Health Initiative
- European Commission
- Horizon 2020 Framework Programme