Quantitation of Bacteroides forsythus in subgingival plaque

Shelburne, Charles E.; Prabhu, Anila; Gleason, Raymond M.; Mullally, Brian; Coulter, Wilson A.

doi:10.1016/s0167-7012(99)00106-2

Cited by 65 publications

(11 citation statements)

References 43 publications

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“…The detection of Aggregatibacter actinomycetemcomitans (previously Actinobacillus actinomycetemcomitans ), Campylobacter rectus , Fusobacterium nucleatum , Prevotella intermedia , Porphyromonas gingivalis , Tannerella forsythia (previously T. forsythensis ), and Treponema denticola in pooled plaque samples was evaluated by real-time qPCR, as described, 20,21 using primers specific for the hypervariable segments of the 16S rRNA genes of each bacterium (Table 1). The percentage of the total flora for each species was calculated by dividing the number of target organisms by the total number of bacteria as determined by qPCR using 16S rRNA primers that reacted with all bacterial species.…”

Section: Methodsmentioning

confidence: 99%

Identification of Pathogen and Host‐Response Markers Correlated With Periodontal Disease

Ramseier

Kinney

Herr

et al. 2009

Journal of Periodontology

314

290

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Background Periodontitis is the major cause of tooth loss in adults and is linked to systemic illnesses, such as cardiovascular disease and stroke. The development of rapid point-of-care (POC) chair side diagnostics has the potential for the early detection of periodontal infection and progression to identify incipient disease and reduce health care costs. However, validation of effective diagnostics requires the identification and verification of biomarkers correlated with disease progression. This clinical study sought to determine the ability of putative host- and microbially derived biomarkers to identify periodontal disease status from whole saliva and plaque biofilm. Methods One hundred human subjects were equally recruited into a healthy/gingivitis group or a periodontitis population. Whole saliva was collected from all subjects and analyzed using antibody arrays to measure the levels of multiple proinflammatory cytokines and bone resorptive/turnover markers. Results Salivary biomarker data were correlated to comprehensive clinical, radiographic, and microbial plaque biofilm levels measured by quantitative polymerase chain reaction (qPCR) for the generation of models for periodontal disease identification. Significantly elevated levels of matrix metalloproteinase (MMP)-8 and -9 were found in subjects with advanced periodontitis with Random Forest importance scores of 7.1 and 5.1, respectively. The generation of receiver operating characteristic curves demonstrated that permutations of salivary biomarkers and pathogen biofilm values augmented the prediction of disease category. Multiple combinations of salivary biomarkers (especially MMP-8 and-9 and osteoprotegerin) combined with red-complex anaerobic periodontal pathogens (such as Porphyromonas gingivalis or Treponema denticola) provided highly accurate predictions of periodontal disease category. Elevated salivary MMP-8 and T. denticola biofilm levels displayed robust combinatorial characteristics in predicting periodontal disease severity (area under the curve = 0.88; odds ratio = 24.6; 95% confidence interval: 5.2 to 116.5). Conclusions Using qPCR and sensitive immunoassays, we identified host- and bacterially derived biomarkers correlated with periodontal disease. This approach offers significant potential for the discovery of biomarker signatures useful in the development of rapid POC chairside diagnostics for oral and systemic diseases. Studies are ongoing to apply this approach to the longitudinal predictions of disease activity.

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Section: Methodsmentioning

confidence: 99%

Identification of Pathogen and Host‐Response Markers Correlated With Periodontal Disease

Ramseier

Kinney

Herr

et al. 2009

Journal of Periodontology

314

290

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show abstract

“…Quantitative real-time PCR was performed to detect and quantify six specific bacteria ( Porphyromonas gingivalis , A ggregatibacter actinomycetemcomitans , Tannerella forsythia , Treponema denticola , Prevotella intermedia , Parvimonas micra ) using species-specific primers [22, 23]. SYBR Green (Life Technologies, Carlsbad CA, USA) was used as nucleic acid stain.…”

Section: Methodsmentioning

confidence: 99%

Single or repeated antimicrobial photodynamic therapy as adjunct to ultrasonic debridement in residual periodontal pockets: clinical, microbiological, and local biological effects

et al. 2013

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This study aims to assess in residual periodontal pockets the clinical, microbiological, and local biological effects of antimicrobial photodynamic therapy (PDT), delivered after ultrasonic instrumentation either once or twice in a 1-week interval. A single center, three-arm randomized longitudinal study was carried out for 6 months. Twenty-eight systemically healthy patients on periodontal maintenance with residual pockets (pocket depth (PD) ≥5 mm, clinical attachment loss ≥2 mm, and bleeding upon probing (BOP+)) were included. Residual pockets on three teeth, separated from each other by at least two other teeth, served as study sites. After ultrasonic debridement, they were randomly assigned to either PDT delivered twice within 1 week (group A), PDT delivered only once (group B), or sham treatment without activating the laser (group C). Methylene blue was applied with a blunt irrigator tip into the pockets. Sites were irradiated with laser light at a wavelength of 670 nm using a light-diffusing tip introduced into the pocket. Initial PD was 5.9 ± 0.9, 6.3 ± 1.3, and 6.3 ± 1.5 mm in groups A, B, and C, respectively, differences being nonsignificant. PD was significantly reduced in all groups. At month 3, PD was significantly lower in groups A (2.9 ± 1.1 mm; p = 0.04) and B (2.8 ± 1.1 mm; p = 0.03) compared to group C (3.5 ± 1.2 mm). At month 6, none of the sites in group A had persisting pockets PD >4 mm and BOP+, whereas two sites in group B and four sites in group C stayed in this category. Detection frequencies of the studied microorganisms at >1,000 and >100.000 cells/ml did not change significantly from baseline to months 3 or 6 in any group. A significant overall decrease was observed from baseline to month 6 for C-reactive protein, serum amyloid A, fibrinogen, procalcitonin, and α-2 macroglobulin. When looking at the groups separately, C-reactive protein was significantly lower only if the laser had been activated twice (p < 0.05). Other differences between groups were not significant. A single or double episodes of PDT had some additional benefit over ultrasonic instrumentation alone.

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“…Colonization of plaque samples collected simultaneously with the serum in the CP groups were evaluated by real-time PCR as described [37] using primers specific for P. gingivalis (forward:5′-CATAGATATCACG-AGGAACTCCGATT-3′; reverse 5′-AAACTGTTAGCAACTACCGATGTGG-3′) and F. nucleatum (forward: 5′-AAATATGTTGAATTCTGGAAAGAGTTTG-3′; reverse: 5′-TGAACTCCAGCTTTTATACTTCTACCAA-3′). Percentage of the total flora for each species was calculated by dividing the number of target organisms by the total number of bacteria as determined by realtime PCR using 16S rRNA primers that reacted with all bacterial species (forward: 5′-CCATGAAGTCGGAATCGCTAG-3′; reverse: 5′-GCTTGACGGGCGGTGT-3′).…”

Section: Methodsmentioning

confidence: 99%

Serum Antibodies to Porphyromonas gingivalis Chaperone HtpG Predict Health in Periodontitis Susceptible Patients

Shelburne

Dhople

et al. 2008

PLoS ONE

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BackgroundChaperones are ubiquitous conserved proteins critical in stabilization of new proteins, repair/removal of defective proteins and immunodominant antigens in innate and adaptive immunity. Periodontal disease is a chronic inflammatory infection associated with infection by Porphyromonas gingivalis that culminates in the destruction of the supporting structures of the teeth. We previously reported studies of serum antibodies reactive with the human chaperone Hsp90 in gingivitis, a reversible form of gingival disease confined to the oral soft tissues. In those studies, antibodies were at their highest levels in subjects with the best oral health. We hypothesized that antibodies to the HSP90 homologue of P. gingivalis (HtpG) might be associated with protection/resistance against destructive periodontitis.Methodology/Principal FindingsELISA assays using cloned HtpG and peptide antigens confirmed gingivitis subjects colonized with P. gingivalis had higher serum levels of anti-HtpG and, concomitantly, lower levels of attachment loss. Additionally, serum antibody levels to P. gingivalis HtpG protein were higher in healthy subjects compared to patients with either chronic or aggressive periodontitis. We found a negative association between tooth attachment loss and anti-P. gingivalis HtpG (p = 0.043) but not anti-Fusobacterium nucleatum (an oral opportunistic commensal) HtpG levels. Furthermore, response to periodontal therapy was more successful in subjects having higher levels of anti-P. gingivalis HtpG before treatment (p = 0.018). There was no similar relationship to anti-F. nucleatum HtpG levels. Similar results were obtained when these experiments were repeated with a synthetic peptide of a region of P. gingivalis HtpG.Conclusions/SignificanceOur results suggest: 1) anti-P. gingivalis HtpG antibodies are protective and therefore predict health periodontitis-susceptable patients; 2) may augment the host defence to periodontitis and 3) a unique peptide of P. gingivalis HtpG offers significant potential as an effective diagnostic target and vaccine candidate. These results are compatible with a novel immune control mechanism unrelated to direct binding of bacteria.

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Quantitation of Bacteroides forsythus in subgingival plaque

Cited by 65 publications

References 43 publications

Identification of Pathogen and Host‐Response Markers Correlated With Periodontal Disease

Identification of Pathogen and Host‐Response Markers Correlated With Periodontal Disease

Single or repeated antimicrobial photodynamic therapy as adjunct to ultrasonic debridement in residual periodontal pockets: clinical, microbiological, and local biological effects

Serum Antibodies to Porphyromonas gingivalis Chaperone HtpG Predict Health in Periodontitis Susceptible Patients

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