Toxoplasmosis has been described as the most widespread zoonotic disease of humans and other animals. Although Toxoplasma gondii infection in high school girls has been investigated in many countries, surveys have not been available in high school girls in Yemen. This study was performed to investigate the seroprevalence and risk factors associated with T. gondii infection among high school girls in Ibb city, Yemen. The study was carried out during the period February to December 2016. The sera from 220 volunteer girls were evaluated for T. gondii antibodies (IgM and IgG) using ELISA method. The seropositive rate of T. gondii was 18.2%, and 81.8% of high school girls was seronegative in anti-Toxoplasma IgG, while all cases (100%) were seronegative for IgM. Among the risk factors evaluated, only the presence of cats in home (p = 0.004) was observed as a significant risk factor associated with T. gondii infection. The results of this study may be useful for the design of optimal preventive measures against infection with T. gondii.