Background: Prenatal screening offers crucial information about the pregnancy, although sometimes brings forth ethical issues and potentially difficult decisions for pregnant women. Ultrasound screening during pregnancy is both of particular importance and a great challenge. Objective: The objective of this study is to assess women’s knowledge and expectations regarding prenatal ultrasound screening in Greece. Methods: The study was conducted between May 2019 and January 2020. Three hundred pregnant women >18 years of age with complete antenatal routine screening, were invited to participate. The questionnaire contained sociodemographic and obstetric history information and highlighted the background knowledge of pregnant women in prenatal screening. Results: In this study, women with university or technological education mentioned in a greater proportion the importance of prenatal screening regarding prevention and treatment of intrauterine diseases. The belief that the pain or sorrow to give birth to a disabled child or one with a genetic disorder might lead parents to the decision to terminate the pregnancy, was supported more by women with a higher educational level. Conclusion: Pregnant women place a great deal of trust in their physician, as a source of information for the fetal well-being. The majority consider prenatal testing mandatory for that matter. Finally, an association between women who abide with prenatal screening and their educational level was noted. Thus, it’s more common women who have a higher educational level to seek prenatal screening, than those with lower education level.