M Yasin Tipu and Zafar Iqbal Ch Bovine tuberculosis is not only a threat to animal but is also a zoonotic and threat to public health. Mycobacterium bovis, the cause of bovine tuberculosis in humans (extra-pulmonary tuberculosis, usually of throat glands in rural population) and M tuberculosis (the cause of classical lungs tuberculosis in humans). Both of the species of TB causing organisms are genetically and antigenically similar. The disease is a major problem and is causing more causalities worldwide than even malaria and HIV. TB is present worldwide with an extremely high prevalence is Asian countries, where 60-80% of children below the age of 14 years are infected. The largest number of cases occurs in the South-East Asia Region, which accounts for 33 % of incident cases globally. Bovine type may cause as high as 25.8% of TB in Man. M bovis has been reported to be cause of 50% of cervical lymph adenitis cases in children in Europe. M bovis is the major cause of human gastrointestinal tuberculosis in developing countries where bovine milk had often not been pasteurized before use. In Pakistan, tuberculosis is prevalent at epidemic proportions in animals and humans. According to an estimate, Pakistan has a population of about 34.3 million cattle and 30.8 million heads of buffaloes. Among the diverse bovine pathogens, Mycobacteria are highly significant, especially for farmed animals. Apart from mortality, it is estimated that animal affected with tuberculosis lose 10-25% of their productive efficiency. Bovine TB has never been seriously attended. Practically this disease and its various aspects that are required to be attended still remain marginalized. It is hypothesized that the bovine T.B. may be prevalent in Pakistan population due to use of raw un-boiled milk. The existing situation calls for a comprehensive program to address this problem in order to save livestock and particularly the human population at risk. Key factors in the control of tuberculosis are rapid detection, adequate therapy, and contact tracing to arrest further transmission. Bovine tuberculosis infection in cattle is usually diagnosed in the live animal on the basis of delayed hypersensitivity reactions. After death, it is diagnosed by post-mortem examination and histopathological and bacteriological techniques. Bacteriological examinations may comprise the demonstration of acid-fast bacilli by microscopic examination (provides presumptive confirmation), and the isolation of mycobacteria on selective culture media and their subsequent identification by cultural and biochemical tests. Culture is gold standard, but a lot of time is required for this slow-growing organism. Serodiagnosis of tuberculosis also has many limitations. As an alternative to these conventional methods, nucleic acid based techniques show promise as more rapid, sensitive and specific means of identification of mycobacteria with some limitations. There was need to develop a PCR-based method as an inexpensive, efficient, single step and sensitive multiplex PCR from three different well established DNA target amplification and to use this newly developed multiplex PCR system successfully for differentiation of M. tuberculosis complex in cattle/ buffaloes and humans, especially in developing countries, like Pakistan. In a study held at Department of Pathology, University of Veterinary and Animal Sciences Lahore, attempts have been made to check the surveillance and standardize Polymerase Chain Reaction (PCR) based techniques for the detection of TB in buffaloes, cattle and animal products (milk and meat). An evaluation and comparison was made between the modern (PCR) and conventional methods (Tuberculin Test, isolation and identification by cultural examination and histopathological studies) for rapid diagnosis of the Mycobacterial spp. The study was performed in different experiments involving Buffaloes, Cattle and animal food products. It has been observed that PCR had more sensitivity and required less time to detect and differentiate different Mycobacterial species as compared to conventional methods. It was also noted that M bovis were found in milk and blood of milking animals as well as some tissue samples collected from Lahore slaughter house. On the basis of findings, the regular monitoring of the milking animals, milk men and animals is suggested. A major contributor in the spread of infection is the sale of body parts of slaughtered in the slaughter houses which are being sold in large cities, including Lahore for the purpose of Sadka.