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Background: Vitamin D plays a very important role in the human body. In Saudi Arabia, the sun exposure is limited for many reasons. The use of vitamin D-fortified food or supplements may complement the daily body requirements. The knowledge and perception of the general population towards vitamin D-fortified intake in Jazan, Saudi Arabia, are not known. Objectives: In this study, we aim to evaluate the knowledge and attitude toward vitamin Dfortified food intake among the general population in the Jazan region. Methods: We performed a cross-sectional questionnaire-based study using the convenient sampling method on the Jazan general population. The questionnaire was composed of questions distributed over three domains. The data were analyzed using SPSS software version 23. Frequency and percentages were used to display categorical variables. Mean and standard deviation were used to present numerical variables. The independent t-test and analysis of variance (ANOVA) test were both used to test for factors associated with knowledge and perception score. Results: A total of 921 participants were included in the study. The majority of participants (72.2%) were between 20 and 40 years; the female gender and Saudi nationality represent the majority of participants in this study. In terms of occupation, 32.9% were currently employed, and 44.5% were students. Regarding education, 64.9% had a bachelor’s degree, and 19.9% had a high school education. The minimum knowledge score was 0, the maximum was 25, and the mean was 10.6 + 4.35. The knowledge levels toward the vitamin D-fortified food intake showed that 64.9% had a moderate knowledge level. With regard to the assessment of participants’ perception towards vitamin D-fortified food intake, 69.6% reported having a vitamin D-rich food before, while 5.8% reported they never did, and 34.6% did not know. The most commonly reported reason for never having vitamin D-fortified food was not knowing what food to take for 19 participants (35.8%). Occupation and education levels of participants were significantly associated with knowledge of vitamin D-fortified food intake. Conclusion: Despite the population in the current study having a moderate level of knowledge about vitamin D-fortified food, the practices towards intake of vitamin D-fortified food or supplements are still underused.