The additional value of real‐time PCR in the quantitative detection of periodontal pathogens

Boutaga, Khalil; Winkelhoff, A.J. van; Vandenbroucke-Grauls, Christina M. J. E.

doi:10.1111/j.1600-051x.2006.00925.x

Cited by 89 publications

(117 citation statements)

References 33 publications

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“…In fact, all the strains which were not detected with the Invader assay had counts lower than 10 5 in real-time PCR. However, levels of periodontopathic bacterial species in subgingival plaque and saliva of patients with periodontitis ranged from 10 4 -10 7 copies (11,(21)(22)(23). These results suggest that the Invader assay is capable of detecting levels of periodontopathogens in patients with clinically problematic periodontal conditions.…”

Section: Discussionmentioning

confidence: 92%

Quantification of Periodontopathic Bacteria in Saliva Using the Invader Assay

et al. 2012

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SUMMARY: When quantifying periodontopathic bacteria, it is important to use a convenient method that does not produce false negative results. The Invader assay is a convenient method because it does not involve gene amplification. The purpose of this study was to evaluate the validity of the Invader assay to quantify periodontopathic bacteria. The Invader technology was applied in quantifying five periodontopathic bacteria (Aggregatibacter actinomycetemcomitans, Porphyromonas gingivalis, Tannerella forsythia, Prevotella intermedia, and Treponema denticola). The Invader assay produced a linear quantitative detection range over concentrations spanning seven exponential values, with a detection limit of 10 3.7 copies/tube and intra-day and inter-day variance of 0.1z to 4.7z and 0.1z to 3.4z, respectively, in quantifying five periodontopathic bacteria. We compared the results of the Invader assay with those of real-time polymerase chain reaction (PCR) performed for quantifying five periodontopathic bacteria in 22 patients with periodontitis. Among the Invader-detectable bacterial strains of each species, significant correlations were observed in the counts of concerned bacterial species between these two methods, with correlation coefficients ranging from 0.757 to 0.996. This study validated repeatability and reproducibility of the Invader assay in quantifying periodontopathic bacteria and demonstrated consistent agreement between the Invader assay and real-time PCR in quantifying periodontopathic bacteria.

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Section: Discussionmentioning

confidence: 92%

Quantification of Periodontopathic Bacteria in Saliva Using the Invader Assay

et al. 2012

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“…The primers and probes selected for the following bacteria were as published in the sources shown: P.gingivalis A. actinomycetemcomitans, P. intermedia, P. micra, T. forsythia, F. nucleatum [29][30][31][32] T. denticola [33], S. mutans, F. nucleatum, S. oralis, A. naeslundi [34], V. dispar [35] and were purchased from All reaction efficiencies calculated were acceptable (between 91% and 104%). All primer sets failed to amplify the DNA purified from different microbial standards.…”

Section: Real-time Quantitative Polymerase Chain Reactionmentioning

confidence: 99%

Is there an association between obstructive sleep apnea syndrome and periodontal inflammation?

et al. 2015

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Objectives: To assess salivary, serum biomarkers, and subgingival bacteria as putative candidates in the potential association between obstructive sleep apnea syndrome (OSAS) and periodontal disease. Materials and methods:Fifty-two patients were grouped according to the severity of OSAS: 13 participants served as controls, 17 patients had mild-to-moderate OSAS, and 22 severe OSAS. Serum, saliva, and subgingival plaque samples were collected, clinical periodontal parameters recorded. Salivary, serum concentrations of interleukin-6 (IL-6), tumour necrosis factor (TNF-α), osteoprotegerin, sRANKL, and apelin were analysed by enzyme-linked immunosorbent assay. Bacterial counts were determined by real-time QPCR on plaque microbial DNA preparations.Results: There was a significant change in the composition of microbes in plaque particularly in severe OSAS samples (p<0.01). Statistical analyses indicated significantly higher salivary IL-6 levels in both OSAS groups compared to controls (p<0.05). Salivary apelin levels were significantly higher in Serum levels of these biomarkers and salivary osteoprotegerin, sRANKL levels were similar in the study groups. The incidence and duration of apnea positively correlated with clinical periodontal parameters (p<0.05). Conclusion:OSAS appeared to alter the tested bacteria in plaque, correlate with increasing periodontal disease severity, have additive effect on salivary IL-6.Clinical relevance: OSAS is likely to interact with periodontal disease.

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“…Although classic method such as cultivation are used for oral microbial evaluations, but the microbial profile can be significantly underestimated because many bacteria (especially anaerobes) cannot be cultivated through these techniques. Also isolation and cultivation of the oral flora is accepted as the gold standard for many years (18).…”

Section: Introductionmentioning

confidence: 99%

Development and Comparison of Conventional PCR and SYBR Green Real Time PCR for Detection of Aggregatibacter actinomycetemcomitans and Tannerella for Sythensis

Soleimani

Zolfaghari

Morovvati

2013

Jundishapur J Microbiol

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Background: Aggregatibacter actinomycetemcomitans and Tannerella forsythensis are two major pathogens in destructive periodontal disease in humans. The detection of these bacteria is needed for diagnosis and management of the mentioned diseases. Objectives: We aimed to develop and compare improved multiplex conventional and SYBR green real time PCR assays for a specific diagnosis of the organisms based on specific marker genes. Materials and Methods: Both PCR approaches were performed with primers targeting diagnostic genes of the organisms, hbpA gene of A. actinomycetemcomitans and 16S rRNA gene of T. forsythensis. For preparation of a stable positive control, the PCR products were cloned into pTZ57R/T plasmid. The test specificity was evaluated using the same PCR reactions but in the presence of genomes of various negative control bacteria. Results: As expected, agarose gel electrophoresis of PCR products of the hbpA and 16S rRNA genes showed 160 bp and 250bp bands respectively. Temperature melting analyses of the SYBR green real time PCR assays showed the Tm at 78.02 °C and 84.62 °C for hbpA and 16S rRNA genes respectively. The amplification results using negative control genomes as template was negative showing the specificity of both designed assays. The detection limits of the conventional PCR assay for hbpA and 16S rRNA genes were 5200 and 1200 copies of each gene, respectively. The designed SYBR green PCR assays tenfold increased the test sensitivity. Conclusions: The designed assays provide simple, reliable, and rapid procedures that identify two main periodontal pathogens. Theses assays are new diagnostic opportunities in our laboratory and also are working as important supplements of the current time consuming phenotypic assays.

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The additional value of real‐time PCR in the quantitative detection of periodontal pathogens

Abstract: RT-PCR provides a new rapid diagnostic tool and opens the opportunity to detect small numbers of oral pathogens in clinical specimens, which are under the detection limit by culture technique.

Cited by 89 publications

References 33 publications

Quantification of Periodontopathic Bacteria in Saliva Using the Invader Assay

Quantification of Periodontopathic Bacteria in Saliva Using the Invader Assay

Is there an association between obstructive sleep apnea syndrome and periodontal inflammation?

Development and Comparison of Conventional PCR and SYBR Green Real Time PCR for Detection of Aggregatibacter actinomycetemcomitans and Tannerella for Sythensis

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