INTRODUCTION: Although the worldwide incidence of tuberculosis (TB) is slowly decreasing, annual infection rates in Peru remain among the highest in the Americas. Pharmacies could play an important role in facilitating early detection of TB. However, the awareness, expertise and cooperation of pharmacy workers is fundamental. This study explored the TB-related knowledge, attitudes and practices of pharmacy workers in a district with one of the highest incidences of TB in Peru. METHODS: This cross-sectional study applied a questionnaire that was administered face to face using smartphones with one pharmacy worker at each of 45 randomly selected pharmacies in the El Agustino district of Lima, Peru. RESULTS: Participants were primarily female (78%) and had an average age of 31.3 years old (range 18-57 years old). Only 11% of participants were pharmacists with complete university training. The pharmacy workers' knowledge was adequate; however, workers had important knowledge gaps and myths regarding prevention of TB transmission. Most pharmacy workers (77%) reported they would send a client with a history of cough for more than two weeks to a healthcare center, while 23% reported they would offer them antitussive medication or antibiotics. Almost all workers reported talking with clients about diseases and reported respiratory symptoms as one of the most common causes for consultation (60%). Most participants expressed interest in learning more about TB and expanding their involvement in the fight against TB in their community. CONCLUSION: Pharmacy workers have adequate knowledge about TB. However, we identified gaps in knowledge with respect to prevention of TB transmission. Pharmacy workers commonly see patients with respiratory symptoms and some offer recommendations, including for treatment. Pharmacy workers are willing to learn more and contribute to TB control and could be a valuable asset in the control and prevention of TB in Peru. To achieve this integration of pharmacy workers into TB control and prevention, more research is needed.