Background: Tuberculous pleural effusion accounts for roughly 30% of extrapulmonary tuberculosis cases, and detecting Mycobacterium tuberculosis in these samples is challenging due to the paucibacillary nature of the disease. The World Health Organization has devised the Gene Xpert Assay for rapid detection of tuberculosis and rifampicin resistance within two hours. This study evaluates the sensitivity and specificity of the Gene Xpert Assay in comparison to Ziehl-Neelsen (ZN) staining and fluorescent microscopy. Materials and Methods: A total of 1342 samples were processed during 2018-2019, including 144 pleural fluid samples. Each sample underwent Gene Xpert Assay, ZN staining, and fluorescent microscopy. Sensitivity and specificity were calculated for each method, and rifampicin resistance was assessed. Results: Of the 144 pleural fluid samples, 9 (6.53%) were positive for tuberculosis using the Gene Xpert Assay, and 1 sample (11.1%) showed rifampicin resistance. Slide positivity rates were 60% for ZN staining and 75% for fluorescent staining. The sensitivity rates were 31% for Gene Xpert, 58.2% for ZN staining, and 63% for fluorescent staining. Conclusion: The Gene Xpert Assay demonstrates lower sensitivity compared to ZN staining and fluorescent microscopy for detecting tuberculous pleural effusion. However, it provides rapid detection and rifampicin resistance identification, which is critical for timely and appropriate treatment.