Strains of the bacterium Actinobacillus actinomycetemcomitans found in the human oral cavity are divided into five serotypes, a, b, c, d, and e. In this study, A. actinomycetemcomitans serotypes and Porphyromonas gingivalis were isolated from 656 subgingival sites in systemically healthy Japanese adults. A. actinomycetemcomitans was detected in 19.5% of 328 Japanese subjects, while 27.1% of subjects were positive for P. gingivalis. Of 75 A. actinomycetemcomitans-positive sites, only one serotype was detected in 39 sites (52.0%). The numbers of sites in which two different serotypes and three different serotypes were detected were 18 (25.0%) and 7 (9.3%), respectively. A. actinomycetemcomitans serotype c was detected more frequently in sites that were positive for both P. gingivalis and A. actinomycetemcomitans (76.9%) than in sites that were P. gingivalis-negative and A. actinomycetemcomitans-positive (33.9%). In addition, serotype c was detected much more frequently than the other serotypes (<16%) in sites that were positive for both P. gingivalis and A. actinomycetemcomitans. These findings suggest that the characteristics of serotype c may differ from those of the other serotypes. This report is the first to use PCR to describe the distribution of A. actinomycetemcomitans serotypes in humans and to examine the association between the distribution of A. actinomycetemcomitans serotypes and the presence of P. gingivalis.