Cryptosporidiosis is an opportunistic zoonotic disease considered a global public health concern. Cats are a potential host for transmitting zoonotic diseases to humans as Cryptosporidium spp. infection. The present study aimed to determine the Cryptosporidium spp in stray cats using molecular methods in Al-Qadisiyah governorate, Iraq, from September 2021 to February 2022. A total of 100 fecal samples were collected from different geographical areas. All the samples were investigated using Nested-PCR and sequencing methods to identify and characterize Cryptosporidium spp. in stray cats. The results of the molecular examination showed that 35% of the fecal samples were infected with Cryptosporidium spp. Two species have been identified, C. parvum and C. canis. In conclusion, the results showed that cats are a reservoir host for Cryptosporidium spp for human infection. For the prevention and control of Cryptosporidium spp. Cat infection should get more attention from health officials, especially when the prevalence rate is high.