Introduction: Neural Tube Defects (NTD) remain a major public health burden globally. It is widely acknowledged that folic acid plays a vital role to reduce the risk of developing NTDs. The study aimed to determine knowledge, and practice regarding periconceptional use of folic acid for the prevention of NTD among women in childbearing age at Boralesgamuwa Medical Officer of Health (MOH) area, Sri Lanka.Methods: A descriptive cross-sectional study was carried out among randomly selected 422 women in childbearing age in the said MOH area. Data were collected through a pre-tested interviewer administered questionnaire. The overall knowledge level was classified as adequate (>75%), moderately adequate (50-75%) and inadequate (<50%).Results: Out of 422 participants, the majority was Sinhalese (95.7%), married (66.8%) and unemployed (62.1%). Mean age was 32.02 ±8.625 years. Nearly 56 % had heard about NTDs and 95.5% had heard about folic acid. A majority (279, 66.1%) had ‘inadequate’ level of knowledge, 120 (28.4%) had ‘moderately adequate’ knowledge and only 5.5% (n=23) had ‘adequate’ knowledge. Approximately 60% had taken folic acid and out of them only 42.4% had started folic acid before pregnancy. Age (p<0.001), marital status (p<0.001), parity (p<0.001), educational qualification of participants (p<0.001), monthly household income (p=0.004) and where health services sought from (p <0.001) were associated with the level of knowledge.Conclusions: Majority of participants had inadequate knowledge regarding periconceptional use of folic acid for the prevention of NTDs. However, out of those who have used FA, majority had started to take it before pregnancy. The study findings warrant more educational programmes on the topic bridging the gap of knowledge among women in childbearing age in the study setting.