This study sought to investigate the actual status of awareness regarding the appropriate use of antimicrobial drugs among patients of a wide age range who visit dispensing pharmacies in order to explore more effective intervention methods for improving awareness of the appropriate use of antimicrobial drugs for patients. A questionnaire survey was conducted with 1301 patients who visited different Tsuruha Holdings-operated pharmacies between 1 September 2018 and 31 October 2018. Using multiple regression analysis, we calculated scores based on the patients’ answers regarding their knowledge of antimicrobial drugs and antimicrobial resistance and examined factors related to these scores. Of the 1185 respondents who successfully completed the survey (mean age ± SD, 52.5 ± 18.2 years), 37.2% were 60 years old or older, 13.2% had never or were not sure whether they had taken antimicrobial drugs, and 73.2% did not understand the problem of antimicrobial resistance. Older age, the non-usage of drugs and self-reports of the “lack” of knowledge of antimicrobial resistance were identified as the group that needs education regarding the appropriate use of antimicrobial drugs. Intervention studies should be conducted to examine the efficacy of interventions based on these factors.