Objective: Emergency medical services (EMS) personnel are at a high risk of spreading infections. Compliance with the EMS handwashing regulations is difficult due to frequent dispatches and on-site treatment. This study sought to investigate the education status and awareness about handwashing of EMS personnel, their performance of the recommended handwashing pattern, and their compliance with infection control protocols. Methods: A total of 677 paramedics were educated on handwashing methods and performance and 76 of them were surveyed on handwashing awareness and performance. The survey tool comprised a total of 15 questions, including three on general characteristics, six recognition questions, and six performance questions. The data were analyzed using the SPSS version 17.0 program. Results: Sixty-five percent of the EMS personnel were educated on handwashing methods. The higher the work experience, the higher the performance of handwashing after personal hygiene activities. Hence, there were significant differences in performance among the personnel (P<0.001). Overall, it was found that EMS personnel with a high awareness of handwashing methods, regardless of their occupation, had a good knowledge of the appropriate methods of cleaning and disinfecting ambulances and were able to treat and control infection well. The higher their awareness regarding handwashing, the higher the performance of the same (P<0.001). Conclusion It is necessary to develop annual evaluation programs, a customized education curriculum for each job, and conduct regular education for paramedics.