Bovine tuberculosis is a chronic bacterial disease caused by Mycobacterium bovis. It can be transmitted to humans through direct contact with infected cattle or consumption of unpasteurized milk and milk products. The current study was performed to assess whether raw milk produced in three large scale dairy farms in Central Province of Sri Lanka contain any M. bovis. Raw milk samples were collected from 330 cows representing 230 single intradermal comparative cervical tuberculin test reactors and 100 none-reactors. All milk samples were cultured on Lowenstein Jensen medium with/ without sodium pyruvate for the isolation of Mycobacterium spp., and slopes were observed for the occurrence of growth daily for the first week and then at weekly intervals for 8 weeks. Direct Polymerase Chain Reaction (PCR) was performed simultaneously on all milk samples to detect M. bovis after extracting DNA with a commercial kit. The minimum detection level of M. bovis for PCR in milk was 200 CFU/mL. Only two milk samples from reactive cows were positive for acid fast bacilli. However, their cultures were confirmed as non-tuberculous mycobacteria by PCR. Consequently, all milk samples were confirmed negative for M. bovis according to direct PCR. It was concluded that the milk samples from three large scale dairy farms in Central Province of Sri Lanka did not contain M. bovis.