Toxoplasma gondii (T. gondii) is divided three main clonal lineages designated as type I, II, and III and atypical genotypes were also detected. The distribution of T. gondii genotypes varied from one geographic area to another. This study characterized of T. gondii isolates from pregnant women in Jazan. Genetic analysis of the GRA6-coding fragment was performed for T. gondii genotyping using PCR-RFLP method. The seropositive for Toxoplasma-specific antibodies were determined using ELISA and were 27.9% in pregnant women in Saudi Arabia. Women seropositive for Toxoplasma IgG &IgM (GI=30) and for specific IgG (GII=30) were included. Among pregnant women, 83.3% of GI (women seropositive for IgG and IgM) and 90% of GII (women seropositive for IgG) were asymptomatic and observed clinical symptoms were fever (n=4) and cachexia (n=2) and lymphadenopathy (n=1). GRA6-nested PCR was positive in 8 blood samples (13.3%), 5 of GI & 3 of GII seropositive women. RFLP analysis showed the detection of genotype I in 8 samples with no cases coinciding to pattern of type II or type III.