Two hundred cat fecal samples were collected from various nature of rearing, sexes, ages, sources and health statuses for conventional and molecular investigations of the Toxoplasma gondii parasite. Microscopic fecal examination revealed. that 32 of the 200 cats examined were infected with Toxoplasma gondii oocysts, with an overall infection rate of 16%. Stray cats, females, older than one year, domestic cats, and those suffering from diarrhea recorded the highest rates of infection with the parasite, Infection with the parasite was confirmed using a rapid immunochromatography test to detect the parasite antigen. Molecular identification of Toxoplasma gondii isolates was performed. 32 samples positive were examined using conventional tests, DNA was extracted from fecal samples, molecular identification of Toxoplasma gondii using specific target genes using a specific primer. To detect B1 gene. Polymerase chain reaction results showed that 29 samples positive for the parasite. PCR results revealed that stray, female local cats, older cats and diarrheic cats gave high infection rate. Genetic sequencing results for the B1 gene of the Toxoplasma gondii parasite showed that the samples were sequenced. Only five isolates were found out of the total samples sent for sequencing, and they were registered in GenBank under the serial numbers (LC858431, LC858432, LC858433, LC858434, LC858435). Genetic analysis results for these isolates showed a high similarity between the local isolates that carry the registration number in the global database. The highest similarity rate of 100% was detected with some isolates recorded in the global gene bank