Grading system for periodontitis by analyzing levels of periodontal pathogens in saliva

Abstract

Periodontitis is an infectious disease that is associated with microorganisms that colonize the tooth surface. Clinically, periodontal condition stability reflects dynamic equilibrium between bacterial challenge and host response. Therefore, periodontal pathogen assessment can assist in the early detection of periodontitis. Here we developed a grading system called the periodontal pathogen index (PPI) by analyzing the copy numbers of multiple pathogens both in healthy and chronic periodontitis patients. We collected 170 mouthwash samples (64 periodontally healthy controls and 106 chronic periodontitis patients) and analyzed the salivary 16S rRNA levels of nine pathogens using multiplex, quantitative real-time polymerase chain reaction. Except for Aggregatibacter actinomycetemcomitans, copy numbers of all pathogens were significantly higher in chronic periodontitis patients. We classified the samples based on optimal cut-off values with maximum sensitivity and specificity from receiver operating characteristic curve analyses (AUC = 0.91, 95% CI: 0.87–0.96) into four categories of PPI: Healthy (1–40), Moderate (41–60), At Risk (61–80), and Severe (81–100). PPI scores were significantly higher in all chronic periodontitis patients than in the controls (odds ratio: 31.7, 95% CI: 13.41–61.61) and were associated with age, scaling as well as clinical characteristics including clinical attachment level and plaque index. Our PPI grading system can be clinically useful for the early assessment of pathogenic bacterial burden and follow-up monitoring after periodontitis treatment.

The salivary microbiota in health and disease

“…Moreover, several studies have aimed to differentiate patients with periodontitis from orally healthy controls by means of salivary levels of putative periopathogens. For example, a recently published NGSbased study showed that relative abundance of P. gingivalis could discriminate patients with periodontitis from orally healthy controls with an AUC (area under curve) of 0.80 [42], and a PCR-based study of 9 selected periopathogens reported that it was possible to discriminate the severity of periodontitis based on salivary levels of the bacteria tested [43]. Furthermore, salivary levels of periopathogens have been used in periodontal risk assessment.…”

Comparison of Bacterial Load Parameters in Subgingival Plaque during Peri-implantitis and Periodontitis Using the RT-PCR Method

“…In our research we have provided the estimates of actual bacterial load levels of five periodontopathogen species in subgingival plaque during periodontitis and peri-implantitis pathologies using the RT-PCR method. The main advantages of using the PCR-method in dentistry and periodontology are reasoned by time and cost-efficiency, higher sensitivity compared to other approaches, possibility to reproduce results in the same manner, and relatively easy algorithm for quantification of obtained parameters (24,25). Recently, real-time polymerase chain reaction has been applied for qualitative analysis, thus completely replacing the method of bacterial culture (14).…”

“…U našem istraživanju dali smo procjenu stvarne razine bakterijskog opterećenja za pet vrsta parodontopatogenih mikroorganizama u subgingivnom plaku tijekom parodontitisa i periimplantitisa metodom RT-PCR. Glavne prednosti primjene te metode u dentalnoj medicini i parodontologiji uključuju brzinu i cijenu testa, veću osjetljivost u usporedbi s drugim postupcima, mogućnost reprodukcije rezultata i razmjerno jednostavan algoritam za kvantifikaciju dobivenih parametara (24,25). Nedavno je lančana reakcija polimeraze u stvarnom vremenu primijenjena za kvalitativnu analizu kao zamjenska metoda za bakterijske kulture (14).…”

“…Ali takve rezultate našeg istraživanja treba tumačiti s oprezom jer je u drugom istraživanju o sastavu mikrobioma utvrđeno da su, iako su primijećene varijacije vrsta između zahvaćenih i zdravih mjesta, takve razlike bile manje nego među subjektima (30). Analogne su činjenice opisane u mnogobrojnim dosadašnjim istraživanjima u kojima su individualni sastav oralnog mikrobioma i pridružene promjene bili dominantni čimbenici razvoja patologije i na koje trebaju biti usmjereni daljnji algoritmi liječenja (22,25,26).…”

Detection of association between periodontitis and polymorphisms of IL‐1β + 3954 and TNF‐α ‐863 in the Korean population after controlling for confounding risk factors

“…Mouthwash samples were collected by rinsing the mouth with 12 mL of solution for 30 seconds (E-zen Gargle; JN Pharm, Korea). 28 Genomic DNA was extracted from mouthwash samples using an Exgene™ Clinic SV DNA extraction kit according to the manufacturer's instructions (GeneAll). The DNA concentration was assessed using a NanoDrop spectrophotometer (Thermo Fisher Scientific).…”