<bold>Objective.</bold>To determine factors associated with delays in pulmonary tuberculosis diagnosis and treatment initiation in the city of Cali, Colombia. <bold>Methods.</bold>This was a retrospective cohort study of cases of tuberculosis (TB) reported in the TB control program of Cali between January and December 2016. The information was collected from the databases of the TB control program, individual treatment cards, and clinical histories. The variables considered were sociodemographic factors, clinical factors, substance use, and performance of the health service. <bold>Results.</bold>A total of 623 cases were identified, of which 57.0% were male. The median age was 42 years (interquartile range (IQR): 27–60). The median time from onset of symptoms to TB diagnosis was 57 days (IQR: 21–117), and from onset of symptoms to TB treatment initiation was 72 days (IQR: 35–145). A factor associated with longer time from the onset of symptoms to TB treatment was being a previously treated TB patient (coefficient: 123.8 days, 95% confidence interval (CI): 48.3 to 199.3). In contrast, being incarcerated was a protective factor for earlier TB treatment initiation (coefficient: -57.3 days; 95% CI: -92.4 to -22.3). <bold>Conclusions.</bold>Our results provide important information concerning risk factors that are associated with delays in the diagnosis and treatment of tuberculosis, and that are subject to future interventions. Health insurance program managers must work together with health care providers on issues that include patient care, health promotion, and updating TB protocols and standards.