In order to facilitate objective structured clinical examinations (OSCE), some hospitals recruit laypersons to receive medical education and training to become standardized patients. It is worthwhile to examine whether laypersons can improve their health literacy by taking part in medical education and training, and consequently reduce utilization of medical resources. Purpose: There were two objectives: (1) to investigate the current status of health literacy of standardized patients and laypersons; and (2) to compare the difference of health literacy of standardized patients and laypersons. Methods: This is a cross-sectional study where the study population was standardized patients (SPs) and the laypersons (LPs) who came to see a doctor or who visited the Health Education Center for consultation at National Taiwan University Hospital. Survey was done through the short-form Mandarin Health Literacy Scale (s-MHLS) anonymously. The difference in health literacy between standardized patients and the laypersons was examined. The basic information of the respondents was obtained through questionnaires, including: gender, age, education level, marital status, employment status, income level, residence, health condition, medical condition. Results: For the SPs (n = 85), 67 of them were females (78.82%). For the laypersons (n = 120), 79 of them were females (65.83%). The mean age of the SPs and laypersons was 62.94 and 67.20 years old, respectively. The health literacy, as measured by the s-MHLS of the SPs ranged from 5 to 11, with a mean of 10.20; and that of laypersons ranged from 3 to 11, with a mean of 9.50. T-test showed there were significant difference between these two groups (P value = 0.002). Results of the multivariate regression model, however, did not show a significant difference between the SPs and the laypersons participated in this study (P value = 0.692). Limitations and implication of the study were discussed. Conclusions: We reported that the health literacy of SPs were better than the laypersons, though without statistical significance. It may be beneficial for laypersons to receive training in medical education to become SPs with better health literacy. It also helps SPs to know their own health literacy and reminds them to value their own health while at the same time encouraging more people to get involved in related activities as SPs.