Bovine tuberculosis is a chronic disease primarily of livestock. This study aimed to evaluate serodiagnostic tests for diagnosis of Mycobacterium bovis. Serum samples were collected from tuberculin positive reactors cattle for serological testing using ELISA and lateral flow immuno-chromatographic test. The positive tested animals were exposed for isolation and identification of Mycobacterium bovis infection from tuberculin reactor lesion sites. Twenty five isolates out of 35 (71.4%) were detected from total visible and non-visible lesions, were characterized as Mycobacterium bovis isolates in comparing with serological test (TB antibody rapid test) gave 9 out of 35 (25.71%) and in comparing with serological test (indirect ELISA) gave 11 out of 35 (31.43%) . The sensitivity of ELISA and lateral flow test in diagnosis of bovine tuberculosis can be applied as a complementary test in the farms positive for Tuberculin skin test but not as a basic test.