Purpose: To evaluate the learning effect in standard automated perimetry using SITA strategy, central 24-2 program, possible associated factors and spatial distribution in individuals with no perimetry experience. Methods: A total of 55 healthy subjects were submitted to Humphrey perimetry in two different sessions in one day. Reliability and global indices, and threshold sensitivity at each point were compared between the two examinations. The influence of potential factors (age, gender, and educational level) and the spatial distribution were evaluated regarding alterations between these two examinations. Results: The duration of the test was longer in the first session (median, 5.7 min; interquartile range [IQR], 1.7 min) than in the second (median, 5.3; IQR, 1.1 min) (p=0.002). The median (IQR) of false negative errors was 2% (6%) in the first examination and 0% (6%) in the second (p=0.04). The mean (standard deviation) in the mean deviation (MD) global index was -2.31 (1.86) dB in the first examination and -1.73 (1.69) dB in the second (p=0.07). No association was observed between the change in MD and age (p=0.29), gender (p=0.69) and educational level (p=0.27). The changes in threshold sensitivity were greater at the peripheral points than at the central points (p<0.001). Conclusion: The threshold sensitivity increased in the second examination compared to the first. No factors were associated with this change. The changes in threshold sensitivity were more evident at the peripheral points. ABSTRACT INTRODUCTION