Diagnosis of smear-negative but culture-positive pulmonary tuberculosis in resource-poor settings is difficult. To determine the value of assessing response to a trial of antibiotics in the identification of patients with positive cultures but negative Ziehl-Neelsen (ZN) smears, we compared clinicians' diagnoses with culture in 334 consecutive adults with suspected tuberculosis in rural South Africa; 142 patients (43%) had culture-positive pulmonary tuberculosis. Diagnosis by ZN smear alone was insensitive (61%) but highly specific (94%). Only half of the smear-negative but culture-positive cases were correctly identified by failing to respond to a broad spectrum antibiotic. The remainder responded to therapy and were discharged. Diagnostic sensitivity therefore increased to 80%, but specificity fell to 78%. A more rigorous algorithm may improve diagnosis of smear-negative pulmonary tuberculosis in resource-poor settings.