Objective: To assess the knowledge, attitudes, and practices (KAP) on Zika virus infection among pregnant women in Brunei Darussalam by a cross-sectional survey. Methods: Between February and June 2017, we recruited 234 pregnant women from all government healthcare centres at Brunei-Muara district, using a modified systematic sampling approach. A pre-tested and self-administered questionnaire was used and data analysis was conducted using descriptive statistics and multiple logistic regression analyses. Results: The study participants were mainly Malay (87.2%) and their mean age was 28.0 years. The median knowledge score was 13, out of a possible score of 28. Most participants (92.7%) knew that Zika virus was transmitted by mosquito bites whereas some (34.6%) knew that sexual transmission was also possible. Media (radio, television or newspapers) was the preferred source of updated information on Zika virus, followed by healthcare workers (44.0%), government announcements (43.2%), and social media (38.0%). Pregnant women who were 25 years old or older [Adj. OR=3.62 (95% CI: 1.57, 9.51)], not Malays [Adj. OR=3.32 (95% CI: 1.35, 8.55)], and had an average monthly household income of more than BND $3 000 [Adj. OR=4.06 (95% CI: 1.81, 19.44)] were more likely to score higher for knowledge on Zika virus. The median prevention practice score was 23, out of a possible score of 36. Most participants reported wearing covering clothes (98.3%) and kept their living surroundings clean (99.6%). Most participants (88.0%) agreed that Zika is an important issue in their community. Conclusion: We found a lack of knowledge on Zika virus infection among pregnant women attending government maternal and child healthcare centres in Brunei Darussalam, in particular that Zika virus can be sexually transmitted. Such information could be well disseminated at the healthcare centre level. Health literacy studies should be conducted to understand the facilitators and barriers of KAP on Zika virus infection among pregnant women.