ObjectiveTo investigate the effect of evidence-based nursing course in undergraduate nursing students' critical thinking ability. MethodsA prospective cohort study design was performed in students from two nursing undergraduate classes, who elected evidence-based nursing course as an observation group and students lacked of the course as a control group. The Chinese Version of Critical Thinking Disposition Inventory (CTDI-CV) was used to test critical thinking ability at the beginning and the end of the semester. The scores of CTDI-CV and rank of scores were compared between the two groups using t test and rank sum test. ResultsA total of 88 students were included, of which 41 students were in the observation group and 47 students were in the control group. Compared to the control group, the observation group demonstrated significantly more improvement in the total scores (MD=22.29, 95%CI 13.477 to 31.114), the search for truth (MD=2.81, 95%CI 0.211 to 5.410), open-minded (MD=3.37, 95%CI 1.280 to 5.471), analysis capabilities (MD=3.41, 95%CI 1.351 to 5.481), systematicity (MD=3.39, 95%CI 1.447 to 5.336), confidence critical thinking (MD=2.83, 95%CI 0.512 to 5.152), curiosity (MD=4.53, 95%CI 2.273 to 6.786) and cognitive maturity (MD=1.93, 95%CI -0.502 to 4.377). The level of total scores, the search for truth, analysis capabilities, systematicty, confidence critical thinking, and curiosity in the observation group were significantly improved. ConclusionEvidence-based nursing course learning can improve the undergraduate nursing students' critical thinking skill.