This study aimed to investigate the accuracy of gingival thickness measurement by two methods of clinical evaluation and intraoral ultrasonography. The gingival thickness was measured in the midbuccal area of the right maxillary lateral incisor and first molar teeth in 30 individuals. For clinical measurement, a #15 K-file with rubber stops was vertically inserted 2 mm apical to the gingival margin and the length of the file in the tissue was measured using a digital caliper. Ultrasonographic measurement was performed using an intraoral probe on the gingival surface in the midbuccal area, at the entry point of the file. Statistical analysis was performed by paired t-test, correlation coefficient, and receiver operating characteristic (ROC) curve (α = 0.05). In the anterior region, the mean gingival thicknesses using ultrasonography (1.517 ± 0.293 mm) and clinical evaluation (1.610 ± 0.272 mm) were not significantly different (p= 0.434). In the posterior region, the mean gingival thicknesses were significantly different between ultrasonography (1.372 ± 0.442 mm) and clinical evaluation (1.626 ± 0.310 mm) (p= 0.006). The area under ROC curve values for ultrasonographic measurements in the anterior and posterior regions were 0.681 and 0.597, respectively. The use of ultrasonography with an intraoral probe has acceptable accuracy for the determination of gingival thickness, especially for the anterior regions.