Aim: To characterize the subgingival microbiome in subjects with different periodontal health statuses. Materials and Methods: In this cross‐sectional observational study, subgingival samples were harvested from Spanish subjects with different periodontal health statuses, based on the 2018 Classification of Periodontal and Peri‐Implant Diseases and Conditions. Samples were processed using high‐throughput sequencing technologies (Illumina MiSeq). Taxa differentially abundant were identified using Analysis of Compositions of Microbiomes with Bias Correction (ANCOM‐BC). α‐ and β‐diversity metrics were calculated using q2‐diversity in QIIME2. The analyses were adjusted for age, gender and smoking status. Results: The identified subgingival microbiome showed statistically significant differences among subjects, categorized into periodontal health, gingivitis and stages I‐II and III‐IV periodontitis (p <.05). In patients with severe (stages III‐IV) periodontitis, the genera Filifactor and Fretibacterium were detected 24 times more frequently than in periodontally healthy subjects. Similarly, the genera Porphyromonas, Prevotella and Tannerella were detected four times more frequently (p <.05). The genera Granulicatella, Streptococcus, Paracoccus, Pseudomonas, Haemophilus, Actinobacteria, Bergeyella and Capnocytophaga were significantly associated with healthier periodontal status (p <.05). Conclusions: Significant differences were detected in the subgingival microbiome among periodontal health, gingivitis and stages I‐II or III‐IV periodontitis, suggesting overlapping, yet distinguishable microbial profiles. [ABSTRACT FROM AUTHOR]