Sarcocystis spp. are two-host protozoan parasites belonging to the phylum Apicomplexa. Among different known species of Sarcocystis in cattle, only Sarcocystis hominis is important from the public health viewpoint, because of its zoonotic characteristics. This study presents the first molecular identification of S. hominis in native cattle in central Iran. A sample of diaphragm muscle from a 6-year-old native cow slaughtered at Yazd Slaughterhouse, Yazd, central Iran, was collected in May 2013. DNA extraction was performed, using the salting-out method. DNA purification and precipitation were performed consecutively. The amplicon and digestion results were analyzed using agarose gel electrophoresis. A PCR product with 926 bp in length was obtained after amplification, and 376 bp and 550 bp in length after digestion that identified S. hominis. To the best of our knowledge, this study is the first of its kind to be reported from Iran.