Marie Sylvianne Rabodoarivelo, Eik Hoffmann, Cyril Gaudin, Diana Angélica Aguilar-Ayala, Jordana Galizia, Lindsay Sonnenkalb, Michael Dal Molin, Laura Cioetto-Mazzabò, Giulia Degiacomi, Deborah Recchia, Jan Rybniker, Riccardo Manganelli, Maria Rosalia Pasca, Santiago Ramón-García, Ainhoa Lucía
{"title":"Protocol to quantify bacterial burden in time-kill assays using colony-forming units and most probable number readouts for Mycobacterium tuberculosis.","authors":"Marie Sylvianne Rabodoarivelo, Eik Hoffmann, Cyril Gaudin, Diana Angélica Aguilar-Ayala, Jordana Galizia, Lindsay Sonnenkalb, Michael Dal Molin, Laura Cioetto-Mazzabò, Giulia Degiacomi, Deborah Recchia, Jan Rybniker, Riccardo Manganelli, Maria Rosalia Pasca, Santiago Ramón-García, Ainhoa Lucía","doi":"10.1016/j.xpro.2025.103643","DOIUrl":null,"url":null,"abstract":"<p><p>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>.</p>","PeriodicalId":34214,"journal":{"name":"STAR Protocols","volume":"6 1","pages":"103643"},"PeriodicalIF":1.3000,"publicationDate":"2025-03-10","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"0","resultStr":null,"platform":"Semanticscholar","paperid":null,"PeriodicalName":"STAR Protocols","FirstCategoryId":"1085","ListUrlMain":"https://doi.org/10.1016/j.xpro.2025.103643","RegionNum":0,"RegionCategory":null,"ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q4","JCRName":"BIOCHEMICAL RESEARCH METHODS","Score":null,"Total":0}
引用次数: 0
Abstract
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.1.