According to the knowledge, no prior genetic research had identified the Sarcocystis spp. in the definitive host (dogs) in Nineveh province, Iraq. Therefore, the goal of this work was to detect Sarcocystis species in dog feces targeting 18S rRNA gene amplifications. A total of 63 fecal samples from 63 stray dogs randomly chosen from various parts of Nineveh province, Iraq, between April and September 2023, aged (≥8 months) and sex (male=37, female=26), were collected in sterile plastic containers. Two hundred microliters of saline solution are added to 1 gram of feces for each sample to form a fecal suspension for DNA isolation and then Conventional Polymerase chain reaction (cPCR)is performed. The finding showed that out of 63 fecal samples detected by cPCR technique. 4/63(6.34%) of the samples tested positive in dogs, 3 female 11.53% and 1 male 2.7% respectively. Gel electrophoresis for Sarcocystis spp. DNA, showed that the positive bands were approximately 900 bp in Nineveh province for the first time. This work highlights the value of genetic testing for identifying Sarcocystis species and offers an invaluable diagnostic resource for future epidemiological research and the evaluation of the efficacy of this disease's management strategies.