Ornithobilharzia turkestanicum is one of the Schistosomatidae family that causing cercarial dermatitis in humans, heavy infestations and considerable economic casualties in production of animals like; sheep, goats and cow. The present study were designed to detection of O. turkestanicum in sheep at the molecular level by using 28S rRNA gene. Worms were collected from Maysan province/Southern Iraq. Adult worms were observed in the mesenteric veins of infected sheep and identified as O. turkestanicum by traditional methods. Adult flukes used to extraction of DNA from tissue and applied on conventional PCR program to amplification of DNA. Size of PCR product were analyzed by 1% gel agarose was 1009bp in matching with specific DNA ladder (10000bp). Confirmation of molecular diagnosis of O. turkestanicum by using sequencing technique. The sequencing of the amplified PCR products were 100% identical between the adult blood flukes and Gene Bank. The conclusion is the molecular techniques are more accurate to detection of adult O. turkestanicum in compares with ordinary methods.