Nosocomial tuberculosis (TB) infection is still a problem in many Taiwanese hospitals. The objectives of this study were to explore the intervals before TB diagnosis and isolation at a regional hospital in Taiwan, and to provide useful knowledge to hospitals for the purpose of TB infection control. Methods: From 2002 to 2005, we included a total of 343 patients with culture-positive pulmonary TB in a regional hospital in Southern Taiwan for this study. Their medical records were reviewed, and the time intervals between patient-hospital contact points and isolation were recorded. Results: Of 343 culture-positive pulmonary TB patients, the majority were male, over 40 years old, and unemployed. The mean interval between the first admission and isolation was 20.5 days (median, 2.0 days). The mean intervals between the first admission from outpatient clinics, emergency department and hospitalization and suspected TB were < 1 day, 6.07 days and 25.53 days, respectively. The mean accumulated exposure time was 0.35 days, 0.61 days and 10.09 days in outpatient clinics, the emergency department and hospitalization, respectively; 75.5% of patients had their diagnosis confirmed at the chest department of the department of internal medicine. Conclusion: Delayed diagnosis was most likely in the case of hospitalized patients and least likely in outpatient clinics. Delayed diagnosis in hospitalized patients also contributed more severely to TB exposure time than others. Enhancing the quality, speed and ability of specialists and physicians to diagnose TB, especially in emergency departments and in hospitalized patients, is essential.