The aim of this study was to determine the nationwide prevalence of smear-positive tuberculosis (TB) in Bangladesh. A multi-stage cluster survey of a random sample of persons aged =15 years was included in 40 clusters (20 urban, 20 rural). Two sputum samples were collected from study participants and tested initially by fluorescence microscopy and confirmed by the Ziehl–Neelsen method. The crude and adjusted prevalence rates and 95% confidence intervals (CIs) were calculated using standard methods. A total of 33 new smear-positive TB cases were detected among 52 098 individuals who participated in the study. The average participation rate was over 80%. The overall crude prevalence of new smear-positive TB in persons aged =15 years was estimated as 63·3/100 000 (95% CI 43·6–88·9) and the adjusted prevalence was 79·4/100 000 (95% CI 47·1–133·8). TB prevalence was higher in males (n=24) and in rural areas (n=20). The prevalence was highest in the 55–64 years age group (201/100 000) and lowest in 15–24 years age group (43·0/100 000). The prevalence was higher in persons with no education (138·6/100 000, 95% CI 78·4–245·0). The overall prevalence of smear-positive TB was significantly lower than the prevalence estimate of the previous nationwide survey in Bangladesh in 1987–1988 (870/100 000).