In this study, 6150 animals (3600 dairy cattle and 2550 buffaloes) in different Egyptian governorates were tested by single intradermal cervical tuberculin test using PPD-B, 72 cattle (2%) and 26 buffaloes (1%) reacted positively. After slaughtering the PM examination revealed 49 (68.1%) of the slaughtered cattle showed visible lesions (VL); comparing with 17 (65.4%) in buffaloes, while 23 (31.9%) showed non visible lesions (NVL) comparing with 9 (64.6%) in buffaloes. The bacteriological examination of processed samples from the 72 slaughtered cattle
revealed 44 M. bovis isolates (61.1%); 40 (81.6%) were from 49 VL and 4 (17.4%) were from 23 NVL, compared to 15 (57.7%) Mycobacterial isolates from buffaloes; 10 (38.5%) were M .bovis, all were from the 17 VL (58.8%), and 5 isolates (19.2%) were MOTT, 3 (17.6%) were from the 17 VL and 2 (22.2%) were from the 9 NVL. The ESAT6-CFP10 mixture using ELISA of the tuberculin positive animals sera showed VL could detect 83.7% and 70.6% in cattle and buffaloes, respectively; compared to 89.8% and 76.5% in cattle and buffaloes, respectively using PPD-B. On the other hand, the sera collected from tuberculin positive animals with NVL, the antigen mixture revealed 17.4% and 22.2%in cattle and buffaloes, respectively; compared to 21.7% in cattle and 33.3% in buffaloes with the PPD-B. The PCR assay using Oligonucleotide primer that amplifies a 350bp fragment in RD7 region of M. bovis confirmed the cultural and biochemical identification of M. bovis isolates (PCR-positive) and MOTT (PCR-negative