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Molecular Survey of Theileria annulata in Cattle by PCR - RFLP Method in Iran

Jamal Akbari, Javad Javanbakht, Moussa Tavassouli, Mohammad Tabatabai and Rahmehr Shafiei

Theileriosis is one of the infectious diseases of cattle and buffalo in tropical countries and Mediterranean region that is of a considerable economical value, generated by Theileria haemoprotozoan and transferred by vetors belonging to Ixodidea. The present study investigates 52 samples in west and east Azerbaijan, Sanandaj and Kermanshah in Iran (2009). In this study, Theileria annulata was separated from ill and healthy cattle by PCR-RFLP method. Especial primers were achieved from SmI-2 gene for this procedure. A fragment of 270bp was separated from the gathered blood samples. PCR products were distinguished by agarose gel and ultraviolet rays system. Among 22 naturally infected cattle, 13 cases (59.09%) were positive in blood smear and 18 (81.81%) were positive in PCR. Among the random sampled healthy cattle, from 30 cases, 4 cases (13.32%) were positive in blood smear and 5 (61.66%) were positive in PCR. In order to investigate the genetic variation (PCR-RFLP), the separated 270bp fragment was digested by restriction enzymes such as TaqI, BSURT, AIUI, RsaI. TaqI enzyme generatd (200, 170bp) fragments, AIUI enzyme generated (120, 90, 60bp) & (150, 90, 30bp) fragments and RsaI & BSURI enzymes could not digest the considered fragment. Finally, six different patterns in genetic variation of relevant gen were observed.