Objectives:The successful implementation of evidence-based practice (EBP) can lead to appropriate and effective midwifery care during pregnancy, childbirth and in the postnatal period. However, levels of knowledge and confidence in one's ability to apply EBP are related to its effective implementation. This study aimed to investigate levels of knowledge, practice of and self-efficacy towards the use of EBP among midwives in East Iran. Methods: This cross-sectional study took place between January and February 2016 and involved 98 midwives employed at two hospitals and all four urban health care centres in Torbat-e Heydariyeh, Iran. Two subscales of the Evidence-Based Practice Questionnaire were used to assess participants' knowledge and practice of EBP, respectively, while a modified version of a previously described scale was used to determine self-efficacy. Results: A total of 76 midwives participated in the study (response rate: 77.6%). Mean knowledge, practice and self-efficacy scores were 4.48 ± 0.94, 3.53 ± 0.68 and 2.80 ± 0.81, respectively. Significant relationships were found between mean selfefficacy, practice and knowledge scores and proficiency in English language (P = 0.001 each) and statistical methods (P <0.050 each). Additionally, significant relationships were found between knowledge and practice of EBP and proficiency in the use of databases (P <0.050 each). Knowledge and self-efficacy scores were significantly correlated with practice (P = 0.001 each). Conclusion: These findings demonstrate a need for improvement in the self-efficacy, practice and knowledge of EBP among midwives in East Iran. Interventions that promote these factors may help increase the use of EBP in this population.