Aim: The study aimed to investigate the presence of the specific B1 gene T gondii in blood and milk samples from natural infected cattle and pregnant women (16-30 weeks) whose examination performed by the officers at the women's and children's Educational hospital in Al-Diwaniyah, Iraq. Materials and methods: A total of 150 serum samplings were collected analysed and scanned for Anti-T gondi antibodies (75 naturally-infected goats and 75 pregnant women with Toxoplasma). Polymerase chain reaction (PCR) was used to detect of B1(399pb) gene in 26 goat's blood samples and 7 samples from pregnant women. Results: A quick-test anti-cassette gondii results showed 26 positive samples of goats in a percentage of 34,666 percent, while a higher percentage of anti-T prevalence of 7 positive samples in women was 9,333 percent in age groups. T gondii antibodies in sera of goats were 56% in group 2-3 years in sera of goats, while in groups (20-25) years in women were 16%. Also, the higher prevalence of antibodies in goats and pregnant women was also 50% and 57.14% of IgG respectively. PCR results showed that B1gene (399 bp) was detected in 33 blood samples (26 samples of goats and 7 samples from pregnant women). The present of B1 gene in 26 goat's blood samples was 38.46% and 57.14% of the 7 pregnant women's blood samples. Conclusion: In total, 150 milk samples from the same group of goats and pregnant women have been obtained and analysed to investigate T gondii DNA. The amplified B1 gene was detected in 6 milk samples of 75 Goat samples (8%), whereas the findings showed that 75 milk samples of pregnant women did not contain the B1gene.