This guide has been developed to provide practical information on the interpretation of the most commonly used first- and second-line line probe assays (LPA). In addition to the interpretation of the mutations identified by the two assays, this guide includes practical recommendations on additional follow up diagnostic actions that should be performed in the presence of specific mutations and the clinical implications that these mutations have on the selection of appropriate tuberculosis treatment regimens. Additionally, the guide further illustrates individual case studies to demonstrate how information should be reported to clinicians, as well as examples of LPA results, and a recommended reporting template.