Highly Specific Protease-Based Approach for Detection of Porphyromonas gingivalis in Diagnosis of Periodontitis
Open accessAbstract
Porphyromonas gingivalis is associated with the development of periodontitis. Here we describe the development of a highly specific protease-based diagnostic method for the detection of P. gingivalis in gingival crevicular fluid. Screening of a proteolytic peptide substrate library, including fluorogenic dipeptides that contain d-amino acids, led to the discovery of five P. gingivalis-specific substrates. Due to the presence of lysine and arginine residues in these substrates, it was hypothesized that the cleavage was mediated by the gingipains, a group of P. gingivalis-specific proteases. This hypothesis was confirmed by the observation that P. gingivalis gingipain knockout strains demonstrated clearly impaired substrate cleavage efficacy. Further, proteolytic activity on the substrates was increased by the addition of the gingipain stimulators dithiothreitol and l-cysteine and decreased by the inhibitors leupeptin and N-ethylmaleimide. Screening of saliva and gingival crevicular fluid of periodontitis patients and healthy controls showed the potential of the substrates to diagnose the presence of P. gingivalis proteases. By using paper points, a sensitivity of approximately 10(5) CFU/ml was achieved. P. gingivalis-reactive substrates fully composed of l-amino acids and Bz-l-Arg-NHPhNO(2) showed a relatively low specificity (44 to 85%). However, the five P. gingivalis-specific substrates that each contained a single d-amino acid showed high specificity (96 to 100%). This obs…
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Compared with conventional qPCR assays, the overall detection time of the PDA-based sensing platform is reduced to approximately 20 min.
“Compared with conventional qPCR assays, the overall detection time is reduced to approximately 20 min, highlighting its potential for rapid periodontal pathogen screening.75,90”
From Polydopamine-Based Biomaterials in Oral Medicine: Applications, Mechanisms, and Translational Challenges (2026) · cites this paper as [75]
The 16S rRNA of Porphyromonas gingivalis contains GC-rich hairpin regions, which provide suitable targets for nucleic acid-based sensing.
“The 16S rRNA of Porphyromonas gingivalis contains GC-rich hairpin regions, which provide suitable targets for nucleic acid–based sensing.”
From Polydopamine-Based Biomaterials in Oral Medicine: Applications, Mechanisms, and Translational Challenges (2026) · cites this paper as [75]
When evaluated in an artificial saliva matrix, the PDA-based sensing platform for P. gingivalis exhibits a dynamic detection range of 10^2-10^7 CFU mL−1, a limit of detection of 38 CFU mL−1, and recovery rates of 93% to 105%.
“When evaluated in an artificial saliva matrix, this sensing platform exhibits a dynamic detection range of 102–107 CFU mL−1, a limit of detection (LOD) of 38 CFU mL−1, and recovery rates ranging from 93% to 105%.”
From Polydopamine-Based Biomaterials in Oral Medicine: Applications, Mechanisms, and Translational Challenges (2026) · cites this paper as [75]
In vitro antibacterial assays, simplified biofilm models, release studies, and selected animal models show that PDA-based systems can achieve biofilm inhibition, ROS regulation, localized drug release, photothermal/photodynamic antibacterial effects, and inflammation control.
“In vitro antibacterial assays, simplified biofilm models, release studies, and selected animal models show biofilm inhibition, ROS regulation, localized drug release, photothermal/photodynamic antibacterial effects, and inflammation control.”
From Polydopamine-Based Biomaterials in Oral Medicine: Applications, Mechanisms, and Translational Challenges (2026) · cites this paper as [75]
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Aptamer Based SPREETA Sensor for the Detection of <i>Porphyromonas gingivalis</i> G-Protein
Park, S.-G., Lee, H. J., Ji, T., Kim, K., & Ohk, S.-H. (2024) · Journal of Microbiology and Biotechnology · doi:10.4014/jmb.2310.10042 · Cited by 2 · see citing works
Yang, Z., Song, W., Lv, L., Zhai, S., Liu, Y., Cui, H., Li, X., Gao, F., & Chu, S. (2026) · International Journal of Nanomedicine · doi:10.2147/ijn.s611922
“75,90 In addition to pathogen detection, PDA-based platforms have demonstrated excellent performance in the multiplexed analysis of oral cancer–related biomarkers.”
- Authors
- Kaman, W. E., Galassi, F., de Soet, J. J., Bizzarro, S., Loos, B. G., Veerman, E. C. I., van Belkum, A., Hays, J. P., & Bikker, F. J.
- Affiliations
- Erasmus MC (NL)
- Academic Center for Dentistry Amsterdam (NL)
- Department of Preventive Dentistry, Academic Centre for Dentistry Amsterdam, University of Amsterdam, Amsterdam, The Netherlands
- Department of Medical Microbiology and Infectious Diseases, Erasmus MC, Rotterdam, The Netherlands
- Venue
- Journal of Clinical Microbiology
- Publisher
- American Society for Microbiology
- Published
- 2012-01
- Volume
- 50
- Issue
- 1
- Pages
- 104-112
- ISSN
- 0095-1137 (ISSN-L)
- 1098-660X
- PMC
- PMC3256702
- OpenAlex
- W1979016786
- Cited by
- 70
- References
- 0
- Citations by year
- 2026: 5 · 2025: 8 · 2024: 7 · 2023: 8 · 2022: 4 · 2021: 4 · 2020: 3 · 2019: 5 · 2018: 2 · 2017: 3 · 2016: 1 · 2015: 6 · 2014: 5 · 2013: 4 · 2012: 5
- FWCI
- 5.53
- Type
- journal article
- Language
- en
- Open access
- bronze
- doi.org
- Locations
- Journal of Clinical Microbiology (publishedVersion)
- PubMed Central (publishedVersion)
- PubMed (publishedVersion)
- UvA-DARE (University of Amsterdam) (publishedVersion)
- EUR Research Repository (Erasmus University Rotterdam) (publishedVersion)
- VU Research Portal (publishedVersion)
- EUR Research Repository (Erasmus University Rotterdam) (submittedVersion)
- RePub (Erasmus University Rotterdam) (submittedVersion, other-oa)
- Data Archiving and Networked Services (DANS) (submittedVersion)
- Topics
- Health Sciences › Dentistry › Periodontics › Oral microbiology and periodontitis research
- Physical Sciences › Engineering › Biomedical Engineering › Biosensors and Analytical Detection
- Health Sciences › Health Professions › Pharmacy › Oral and gingival health research
- Keywords
- Porphyromonas gingivalis, periodontitis, gingipains, bacterial proteases, gingival crevicular fluid, saliva, protease inhibitors, dithiothreitol, leupeptin, N-ethylmaleimide
- MeSH terms
- Bacteriological Techniques, Gingival Crevicular Fluid, Humans, Peptide Hydrolases, Periodontitis, Sensitivity and Specificity, Porphyromonas gingivalis
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