Oral cancer remains a significant global health problem, with high incidence and mortality rates. Smoking is known to disrupt the balance of the oral microbiota, which has been associated with an increased risk of oral cancer in previous studies. This exploratory, descriptive cross-sectional study aimed to detect the presence of Peptostreptococcus stomatis, Prevotella intermedia, Fusobacterium nucleatum, and Porphyromonas gingivalis, bacterial species previously reported to be associated with tumor microenvironment changes, in oral swab samples from smokers. Swab samples were collected from the soft palate of 9 smokers and 1 non-smoking control, followed by DNA extraction and detection using Real-Time PCR with species-specific primers and SYBR Green. Peptostreptococcus stomatis was the most frequently detected bacterium among smokers, found in 8 of 9 samples (88.9%), followed by Fusobacterium nucleatum (5 samples, 55.6%), Prevotella intermedia (3 samples, 33.3%), and Porphyromonas gingivalis (1 sample, 11.1%); none of the target bacteria were detected in the single non-smoking control. These findings provide a preliminary description of periodontopathogenic bacterial presence in the oral cavity of smokers. Given the small sample size, the single comparator, and the absence of clinically confirmed oral cancer cases, this study cannot establish an association between these bacteria and oral cancer, nor confirm their utility as early-screening biomarkers; larger case-control studies including confirmed oral cancer patients are needed to evaluate this potential.