Bovine tuberculosis is an important and chronic mycobacterial disease of cattle and buffaloes. However very few reports are available on the prevalence of bovine tuberculosis in buffaloes. The present study was thus conducted to detect Mycobacterium tuberculosis complex (MTC) in buffaloes from an organized dairy farm. Comparative intradermal tuberculin test (CITT), Interferon gamma assays (IFN-γ) and Polymerase Chain Reaction (PCR) was used to detect the diseased animals. Out of 188 buffaloes tested by CITT, 17 (9.04%) were positive reactors. Out of these 17 animals, 9 (4.7%) buffaloes were positive by IFN-γ assay. By IS6110 PCR, 17 (9.04%) were positive for MTC. Thus, the study showed that PCR was more sensitive and specific diagnostic technique for tuberculosis. The study provides additional knowledge in the field of tuberculosis in buffaloes and may help in development of strategies for control of the disease.