Background: Tuberculosis (TB), caused by Mycobacterium tuberculosis (MTB), is one of the most common infectious diseases, especially in countries with limited healthcare resources. Early and accurate diagnosis is essential to curb its spread, and loop-mediated isothermal amplification (LAMP) is a rapid, cost-effective, and reliable molecular diagnostic method. Objective: The objective of this study was to evaluate the performance of TB-LAMP as a rapid and reliable diagnostic tool for pulmonary TB and compare it with direct smears and Gene Xpert, using culture as a reference. Materials: Between February and July 2025, a total of 397 patients (227 men and 170 women) were enrolled in the study. Samples were tested by smear microscopy, culture, Xpert MTB/RIF, and TB-LAMP. Results: Of the 397 patients, 84 (21.15%) were culture-positive. Among culture-positive patients, 81 (96.4%) were positive by Gene Xpert, 69 (82.1%) by TB-LAMP, and 49 (58.3%) by direct smear microscopy. Using culture as the reference standard, the specificity, positive predictive value (PPV), and negative predictive value (NPV) of the TB-LAMP assay were 82.1% (73.9%–90.3%), 99.0% (97.9%–100%), 95.8% (91.2%–100%), and 95.4% (93.1%–97.7%), respectively. Conclusions: The TB-LAMP assay exhibits high diagnostic accuracy for the detection of Mycobacterium tuberculosis , making it a rapid, cost-effective, and reliable alternative to conventional methods, especially in resource-poor settings.