Introduction The aims of this study were to confirm the efficacy of the Bolton index in a group of natural Class I white Italian patients with complete dentition, minimal crowding, and no previous orthodontic treatment, and to evaluate the mesiodistal tooth dimensions with multivariate cluster analysis. Methods Tooth measurements were obtained from a sample of 56 patients with normal occlusion, with a normal Bolton index (22 men, 34 women; mean age, 27.8 years), without previous orthodontic treatment or prosthetic or Class II restorations. Maxillary and mandibular measurements were analyzed separately in the sexes. Clustering was performed using the partitioning around medoid algorithm and principal component analysis-based transformed data. Statistical analysis was conducted. Results The multivariate analysis showed 3 distinct clusters of both maxillary and mandibular tooth measurements in the male and female subjects. Statistically significant differences were found between the sexes in terms of average tooth measurements, and there was significant proportionality between the maxillary and mandibular arch clusters. Conclusion Although the Bolton index is useful for identifying dentodental discrepancies in most patients, cluster analysis enabled the mandibular and maxillary tooth dimensions of the male and female subjects to be divided into 3 general classes (clusters) and the precise location of the discrepancies to be pinpointed.