Toxoplasmosis is a zoonotic disease caused by the obligate intracellular protozoan Toxoplasma gondii. The current study assessed the seroepidemiological status and risk factors of T. gondii in the West Bank, Palestine. We conducted a retrospective study among 1,475 patients referred for serological examination for toxoplasmosis between 2017 and 2021 at the Central Public Health Laboratory. The overall prevalences of anti-T. gondii IgG and IgM antibodies were 1.0% [18.9-23.0%] and 1.0% [0.5-1.6%], respectively. Multifactorial analysis showed that age and sex were significantly associated with the presence of IgG antibodies. The prevalence of IgG antibodies was highest in 2018 and decreased significantly each year until 2021. Routine serological screening in women of childbearing age and pregnant women is recommended.