Due to COVID-19 pandemic, there is a large global drop in the number of newly diagnosed cases with tuberculosis (TB) worldwide. Actions to mitigate and reverse the impact of the COVID-19 pandemic on TB are urgently needed. Recent development of TB smear microscopy automation systems using artificial intelligence may increase the sensitivity of TB smear microscopy. The objective is to evaluate the performance of an automation system (μ-Scan 2.0, Wellgen Medical) over manual smear microscopy in a multi-center, double-blind trial. Total of 1726 smears were enrolled. Referee medical technician and culture served as primary and secondary gold standards for result discrepancy. Results showed that, compared to manual microscopy, the μ-Scan 2.0's performance of accuracy, sensitivity and specificity were 95.7% (1651/1726), 87.7% (57/65), and 96.0% (1594/1661), respectively. The negative predictive value was 97.8% at prevalence of 8.2%. Manual smear microscopy remains the primary diagnosis of pulmonary tuberculosis (TB). Use of automation system could achieve higher TB smear sensitivity and laboratory efficiency. It can also serve as a screening tool that complements molecular methods to reduce the total cost for TB diagnosis and control. Furthermore, such automation system is capable of remote access by internet connection and can be deployed in area with limited medical resources.