Salivary Molecular Testing for Periodontal Pathogen Monitoring: Clinical Performance of Flexible RT-PCR Platforms in Preventive Care Settings

<b>Objective:</b> This study aimed to validate the clinical utility of a salivary molecular platform (Oral Predict^®) for periodontal pathogen detection across preventive, therapeutic, and maintenance settings. <b>Methods:</b> A longitudinal randomized study was conducted involving 78 adults who provided saliva samples at baseline, one month, and three months after professional dental hygiene. Participants were randomized into two groups: control group (n = 39) and probiotic group with Oral Predict^® probiotic supplementation (n = 39). Crude saliva was processed directly without nucleic acid extraction and analyzed by multiplex real-time PCR using either the compact Real-time PCR system or standard thermocyclers. <b>Results:</b> At baseline, <i>Fusobacterium nucleatum</i> was the most prevalent pathogen (84.6%), followed by <i>Tannerella forsythia</i> (53.8%) and <i>Porphyromonas</i> gingivalis (46.2%). The Total Pathogen Burden Score (TPBS) showed progressive increases with age, smoking, and poor oral hygiene, and was significantly higher in participants with gingival bleeding. Among individual pathogens, no significant associations were observed with periodontitis staging or grading. Professional hygiene induced mean reductions of 1-2 logs across all pathogens, with TPBS decreasing from 8.7 ± 3.2 to 4.1 ± 2.8 (<i>p</i> < 0.001). At three months, 69.2% of the control group experienced bacterial rebound, whereas 85% of probiotic users sustained or improved bacterial reductions. <b>Conclusions</b>: Salivary molecular testing provides a robust, non-invasive approach for periodontal pathogen detection, treatment monitoring, and long-term maintenance assessment. The flexibility of the Oral Predict^® platform across point-of-care and laboratory settings, combined with automated interpretation, supports integration into preventive protocols and personalized periodontal care. These findings demonstrate the potential of saliva-based molecular diagnostics to shift periodontal management from reactive to predictive and precision-based strategies.