Background: Down syndrome (DS) is the most common chromosomal abnormality found in live-born babies. It is associated with mental retardation, physical disabilities and growth problems. Prenatal screening to identify genetic disorders gaining importance now a days. The aim of this study was to assess the knowledge, attitude and practice among pregnant women about prenatal screening of Down syndrome.Methods: This was a prospective study done in the department of obstetrics and gynecology in Prime Medical Centre, Sharjah. All pregnant women who attended antenatal clinic and gave the consent for the study were included. The study was conducted from April 2019 to September 2019. A self-structured questionnaire was given to these women and data was collected. Responses to pregnant women’s knowledge, attitude and practice were evaluated.Results: A total of 217 pregnant women were included in the study. Of the 217 women 64.97% had good knowledge about Down syndrome but only 23.04% women had good Knowledge regarding Down syndrome screening. Although 42.85% had right attitude towards screening but only 14.74% patients had followed good practice. Expensive test 43.65% and religious beliefs 25.39% were stated as major reasons for denial for screening tests.Conclusions: Pregnant women have good knowledge of Down syndrome but low awareness of screening tests. Although they had positive attitude towards screening test but less patients followed good practice.