Objective: The present study was undertaken to assess familiarity, knowledge, and confidence of medical students with regard to autism spectrum disorders (ASDs). Methods: This study was conducted in a cross-sectional design among medical students in the 3 main universities in Palestine. In addition to the sociodemographic and academic details, the questionnaire measured familiarity (8-items), knowledge (12-items), confidence and willingness to learn (5-items) with regard to ASDs. Results: The questionnaire was completed by309 medical students (response rate = 77.3 %). The median familiarity, knowledge, and confidence scores were 50 % (42.5 %, 57.5 %), 50 % (41.7 %, 66.7 %), and 60.0 % (54.0 %, 68.0 %), respectively. There was a positive moderate correlation between familiarity and knowledge scores (Spearman’s rho = 0.29, p-value < 0.001) and familiarity and confidence scores (Spearman’s rho = 0.34, p-value < 0.001). Medical students who have received a course on autism were 3.08-fold (95 % C.I. of 1.78–5.31) more likely to score ≥ 50 % on the familiarity items compared to those who did not receive a course. Conclusion: The present study identified considerable awareness and knowledge gaps among medical students with regard to ASDs. More studies are still needed to investigate if such interventions can improve healthcare services for individuals with ASDs.