Protocol to quantify bacterial burden in time-kill assays using colony-forming units and most probable number readouts for Mycobacterium tuberculosis

Rabodoarivelo, Marie Sylvianne (Universidad de Zaragoza) ; Hoffmann, Eik ; Gaudin, Cyril ; Aguilar-Ayala, Diana Angélica (Universidad de Zaragoza) ; Galizia, Jordana (Universidad de Zaragoza) ; Sonnenkalb, Lindsay ; Dal Molin, Michael ; Cioetto-Mazzabò, Laura ; Degiacomi, Giulia ; Recchia, Deborah ; Rybniker, Jan ; Manganelli, Riccardo ; Pasca, Maria Rosalia ; Ramón-García, Santiago ; Lucía, Ainhoa (Universidad de Zaragoza)

Resumen: 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.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.

Idioma: Inglés

DOI: 10.1016/j.xpro.2025.103643

Año: 2025

Publicado en: STAR Protocols 6, 1 (2025), 103643 [18 pp.]
ISSN: 2666-1667

Financiación: info:eu-repo/grantAgreement/EC/H2020/853989/EU/EUROPEAN REGIMEN ACCELERATOR FOR TUBERCULOSIS/ERA4TB

Tipo y forma: Artículo (Versión definitiva)

Área (Departamento): Área Microbiología (Dpto. Microb.Ped.Radio.Sal.Pú.)