Introduction and Objectives Clinicians consider musculoskeletal tuberculosis as diagnostic dilemma because diagnosis frequently gets delayed due to its varied clinical presentation, false negative results on smear microscopy and less sensitivity on culture methods (20-40%). The main objective was to know the effectiveness of using ultrasonographic guided aspirate sample in early diagnosis of musculoskeletal tuberculosis. Materials and Methods In this study, a total of thirty seven clinical specimens like pus swab and ultrasonography guided (USG) pus aspiration of newly suspected musculoskeletal tuberculosis patients were evaluated for microscopy, culture and molecular detection of M. tuberculosis. Results The study reveals prevalence of 45.9% musculoskeletal tuberculosis among the clinically suspected patients. The most commonly involved sites in suspected musculoskeletal tuberculosis were hip joint (23.1%). USG guided pus comprises only 26.9% of the samples collected, however, yield of M. tuberculosis from them were 71%. PCR detected the maximum number of cases of musculoskeletal tuberculosis 17 (45.9%) followed by culture method 9 (24%) and smear microscopy 1 (2.7%). Conclusion This observation will help guiding clinicians in effective management of musculoskeletal tuberculosis cases because delayed diagnosis and treatment in musculoskeletal TB results in poor outcome.