The goal of this research was to determine the relationship between the stage of chronic periodontitis and the presence of six bacterial pathogens (Aggregatibacter actinomycetemcomitans: AA, Fusobacterium nucleatum: FN, Porphyromonas gingivalis: PG, Prevotella intermedia: PI, Enterococcus faecalis: EF, and Parvimonas micra: PM). Forty-six chronic periodontitis patients visiting a dental hospital were included in this investigation. They were classified into four chronic periodontitis stages based on the sulcus bleeding index value and the probing depth. The tissue samples from the periodontal surgery were used for a direct PCR detection assay. A total of 49 samples from 46 patients were collected and classified into four chronic periodontitis groups (N: 6, P1: 13, P2: 18, P3: 12). The PCR assay showed that FN, PI, and PM were involved from the beginning of chronic periodontitis (P1), while AA and PG existed regardless of the disease stages. EF was strongly linked to the P3 stage of the disease. In order to assess the effect of dental treatments on patients with chronic periodontitis, EF should be a critical marker for P3 patients, while FN, PI, and PM would be good indicators for chronic periodontitis.