During present study a total of 800 samples were collected from two hundred descriptive (Holstein Friesian) and non-descriptive (Thari) cattle breeds from districts of Mirpurkhas and Badin, 400 samples were taken from each district. Animals were first screened through Single Intradermal Tuberculin Test (SITT) then positive and negative reactors of SITT were further investigated through Rapid BTB Ab test, culture examination and ELISA test. Risk factors such as sex, age, health status, lactating, non lactating and breed were also investigated. In district Mirpurkhas prevalence was 4%, 15%, 2%, 1%, while in district Badin it was 5%, 18%, 3%, 2% through SITT, Rapid BTB Ab test, culture examination and ELISA test, respectively. Prevalence of bovine tuberculosis in Mirpurkhas district via cultural examination was recorded as 1.66%, 2.5% in milk and nasal samples, while in district Badin it was 1.66%, 5%, respectively. The overall prevalence of bovine tuberculosis was 5.5% and 7% in Mirpurkhas and Badin districts through tuberculin, Rapid BTB Ab, Culture examination and ELISA test. Different tests such as SITT, Rapid BTB Ab, culture examination and ELISA test showed that the prevalence of bovine tuberculosis was higher in females, animals above the six years of age, lactating animals, descriptive breed and animals having poor health status. Furthermore, 1% and 2% prevalence was recorded in farm workers at district Mirpurkhas and Badin. It is concluded that disease was prevalent in both districts however; relatively higher prevalence was observed in district Badin.