Objective: The objective of this study was to determine the practice and barriers towards the Pap smear test among Malaysian women.Methodology: This is a cross-sectional survey of female university staff. Data was collected by a self-administered questionnaire from a total of 117 women. T-test and ANOVA test were conducted to determine if there was a significant difference between the study parameters.Results: A total number of 117 female university staff participated in this study. The majority of them were Malays, single, and living in the city (92.3%, 58.1%, and 92.3%; respectively). Regarding lifestyle practices, the majority consumed vitamin and mineral supplements regularly, and exercised once a week, (55.6%, and 41%, respectively). As for their knowledge about cervical cancer, the majority have heard about the Pap smear test before (81.2%). Regarding the practice of Pap smear test, only 22.2% ever had a Pap smear test done. As for the barriers toward having a Pap smear test done, the most common barrier among study participants was lack of time (29.9%), followed by the excuse that the Pap smear test is a painful procedure (17.9%). Regarding the factors that influenced the practice of having a Pap smear test were marital status, occupation, regular vitamin and mineral supplements intake, daily fruits intake, regular medical check-up, educational and income level significantly influenced the practice (p=0.001, p=0.002, p=0.034, p=0.001, p=0.001, p=0.024, p=0.001; respectively).