The trypsin-like peptidase activity assay kit measures the trypsin-like protease produced by three red-complex species, Porphyromonas gingivalis, Tannerella forsythia, and Treponema denticola, causing periodontitis, and detects the presence of these bacteria in samples. The purpose of this study was to investigate the relationship between the detection of TLPs by a novel TLP-AA, ADCHECK and the detection of red-complex pathogens by real-time PCR using tongue swabs from patients with periodontitis. The detection limit of trypsin-like protease activity by ADCHECK was validated using the culture supernatants of two different Porphyromonas gingivalis bacterial strains. Real-time PCR was performed to determine the number of red-complex species in the tongue coatings of patients with periodontal disease. Trypsin-like protease activity in tongue-swab samples was scored using ADCHECK. ADCHECK successfully detected trypsin-like protease activity in 10³ Porphyromonas gingivalis bacterial strains. The specificity, positive predictive value, negative predictive value, and accuracy of ADCHECK for the presence of red-complex pathogens determined by real-time PCR were 90%, 97%, 98%, and 92%, respectively. ADCHECK is an effective tool for the detection of red-complex pathogens.

Periodontitis is a biofilm-driven inflammatory disease in which conventional indices (probing depth, clinical attachment level, and radiographs) quantify tissue destruction without capturing the biology of infection. In this review, we synthesized microbiological diagnostics, from chairside tools to omics. We outline sampling strategies and emphasize the quantitative monitoring of bacterial load. Enzymatic assays (e.g., N-benzoyl-DL-arginine-2-naphthylamide hydrolysis assay test) measure functional activity at the point of care. Immunological methods include rapid immunochromatography for Porphyromonas gingivalis and enzyme-linked immunosorbent assay for the high-throughput measurement of bacterial antigens. Molecular platforms encompass quantitative polymerase chain reaction (qPCR) (TaqMan, SYBR, multiplex panels; propidium monoazide quantitative-qPCR for viable cells), checkerboard DNA–DNA hybridization for semi-quantitative community profiling, loop-mediated isothermal amplification (LAMP)/molecular beacon-LAMP for portable isothermal detection, and microarrays. Complementary modalities such as fluorescent in situ hybridization, next-generation sequencing, and Fourier transform infrared spectroscopy provide spatial, ecological, and biochemical resolutions. We discuss the limitations of current approaches, including sampling bias, presence–activity discordance, semi-quantitation, method biases, limited strain/function resolution, low-biomass artifacts, and lack of validated cutoffs. To address these challenges, we propose a pragmatic hybrid strategy: site-specific quantitative panels combined with activity and host-response markers interpreted alongside clinical metrics under standardized quality assurance/quality control. Priorities include outcome-linked thresholds, strain-aware/functional panels, robust point-of-care chemistry, and harmonized protocols to enable personalized periodontal care.

Porphyromonas gingivalis is the most pathogenic periodontal bacterium in the world. Recently, P. gingivalis has been considered responsible for dysbiosis during the development of periodontitis. This study aimed to evaluate a novel immunochromatographic device using monoclonal antibodies against P. gingivalis in subgingival plaques. A total of 72 patients with chronic periodontitis and 53 periodontally healthy volunteers underwent clinical and microbiological examinations. Subgingival plaque samples were analyzed for the presence of P. gingivalis and compared using real-time polymerase chain reaction (PCR). In the periodontitis group, a significant positive correlation was observed between the test device scores and the real-time PCR results. The specificity, positive predictive value, negative predictive value, and accuracy of the test device for P. gingivalis, as determined by real-time PCR, were 98%, 94%, 89%, and 90%, respectively. There were significant differences in bacterial counts by real-time PCR among the groups with different ranges of device scores. Additionally, there was a significant positive correlation between the device scores for P. gingivalis and periodontal parameters. These results suggest that this novel immunochromatographic device can be effectively used for rapid detection and semi-quantification of P. gingivalis in subgingival plaques.

Background: Periodontal pathogens are related to the incidence of systemic diseases. This study aimed to examine whether periodontal pathogen burden is associated with the risk of fever onset in older adults. Methods: Older adults in nursing homes, aged ≥65 years, were enrolled. The study was set in Kitakyushu, Japan. The body temperatures of participants were ≥37.2 °C and were recorded for eight months. As periodontal pathogens, Porphyromonas gingivalis, Treponema denticola, and Tannerella forsythia were qualified by a real-time polymerase chain reaction at the baseline. For statistical analysis, the number of bacterial counts was logarithmically conversed to 10 as a base. Results: Data from 56 participants with a median age of 88 (62–98) years were available for analysis. The logarithmic-conversed bacterial counts of T. forsythia, but not P. gingivalis or T. denticola, were associated with the onset of fever in older residents. The Kaplan–Meier method revealed that the group with <10⁴ of T. forsythia had significantly less cumulative fever incidence than the group with ≥10⁴ of T. forsythia. The group with ≥10⁴ of T. forsythia was associated with an increased risk of fever onset (hazard ratio, 3.7; 98% confidence interval, 1.3–10.2; p = 0.012), which was adjusted for possible confounders. Conclusions: Bacterial burden of T. forsythia in the oral cavity was associated with the risk of the onset of fever in older nursing homes residents.

Background: Porphyromonas gingivalis is the principal bacterial species implicated in the pathogenesis of periodontitis. Despite its central role, direct detection of periodontopathic bacteria is not routinely performed in dental clinics. Although test kits using monoclonal antibodies against P. gingivalis have been reported, no practical chairside method has yet been established. Objectives: In this study, we developed an immunochromatography system that enables rapid and quantitative detection of P. gingivalis within 10 min. Methods: We evaluated the performance of the developed immunochromatography system and its dedicated measurement reader. Results: The kit demonstrated a minimum detection limit of 2.6 × 10⁴ CFU/mL and maintained detection capability across fimbrial types I and II. The dedicated reader showed strong correlations with a reference reader (R² = 0.9921) and visual color chart evaluation (R² = 0.9347). High intra- and inter-device precision was observed, with coefficients of variation less than 0.6%. Although the presence of 2% whole blood interfered with visual interpretation, other oral substances, including toothpastes and mouthwashes, did not significantly affect the assay performance. Conclusions: Collectively, these findings indicate that the developed system may serve as a rapid and reliable chairside tool for quantitative detection of P. gingivalis, with potential utility in the diagnosis and management of periodontitis and related systemic conditions.