Purpose: Healthcare professionals are encouraged to use evidence-based practice (EBP) to improve the quality of care and the patient’s quality of life. Knowledge and skill set specific to EBP, critical resources such as time and accessibility to databases, and clinician’s attitude toward EBP are needed for successful implementation of EBP. However, little is currently known about these factors among Thai Physical Therapists (PTs). This study aimed to explore the patterns of attitudes, perceived knowledge, and barriers toward EBP in Thai PTs. Method: An online questionnaire was used to obtain information regarding attitudes, knowledge, and barriers from Thai PTs. Descriptive statistics were used to describe the responses. Multiple logistic regression was used to determine the association between personal and work characteristics and the responses related to attitude, knowledge, and barriers. Results: A total of 719 Thai PTs responded to the questionnaire. Based on this sample, Thai PTs had positive attitudes and perceived themselves as having sufficient background EBP education and skills. They were confident in performing a literature search, critically appraisal, and applying research in their practice. However, the frequencies of searching and reading of literature were relatively low. Less than half of respondents understood terms related to EBP asked in the survey instrument. The barriers to EBP were insufficient time and limited research and statistical skills. PTs who were younger and having higher education were more likely to have better attitudes and knowledge. Conclusions: Despite the positive attitude and perceived knowledge of EBP, time allocation, formal training of EBP skills during academic preparation, and continuing education are required to overcome the barriers toward EBP. The discrepancy between the perception and practice of EBP observed calls for further investigation of actual skills and the barriers to the implementation of EBP.