The efficiency of two primer sets that used for diagnosis of Theileria annulata infection in cattle were evaluated in 28 cows that showed clinical signs of theileriosis. Both conventional method and Polymerase Chain Reaction (PCR) were used to confirm theileria infection. For PCR, two primer sets that amplify the 30 kDa major merozoite surface antigen gene were used. Only 12 cows (42.85%) were positive for theileria infection with blood film. Higher number of cows were diagnosed positive by using PCR, the number of positive cows using primer set one (N516/N517) was 21 cows (75%) and by using primer set two (Tams1F/Tspm1R) was 27 (96.42%). It could be concluded that primer set Tams1F/Tspm1R is the first choice when diagnosis of Theileria annulata is decided.