Histograms presenting the age distribution by gender and by group (healthy, caries and periodontitis) for all study participants. While gender is almost equally distributed across all age groups, healthy individuals are more frequently found in younger age groups, caries patients in younger to middle-aged groups, and periodontitis patients in the middle-aged and elderly study population.

Histograms presenting the age distribution by gender and by group (healthy, caries and periodontitis) for all study participants. While gender is almost equally distributed across all age groups, healthy individuals are more frequently found in younger age groups, caries patients in younger to middle-aged groups, and periodontitis patients in the middle-aged and elderly study population.

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Oral health is maintained by a healthy microbiome, which can be monitored by state-of-the art diagnostics. Therefore, this study evaluated the presence and quantity of ten oral disease-associated taxa (P. gingivalis, T. forsythia, T. denticola, F. nucleatum, C. rectus, P. intermedia, A. actinomycetemcomitans, S. mutans, S. sobrinus, oral associated...

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Context 1
... 214 saliva samples were subjected to the final analysis. Due to the explorative nature of this study, gender was not stratified, however, gender was almost evenly distributed over all ages (Figure 2). The age distribution differed slightly between all groups (Figure 2). ...
Context 2
... to the explorative nature of this study, gender was not stratified, however, gender was almost evenly distributed over all ages (Figure 2). The age distribution differed slightly between all groups (Figure 2). Younger subjects frequently belonged to the healthy group (mean: 34 years) and younger to middle-aged subjects to the caries group (mean: 38 years). ...
Context 3
... suffering from periodontitis were mainly found in the middle-aged and elderly population (mean: 54 years). Figure 2. Histograms presenting the age distribution by gender and by group (healthy, caries and periodontitis) for all study participants. ...

Citations

... It presents data on the detection of three caries-associated and seven periodontitis-associated bacterial species in complex saliva samples collected from individuals that were classified into three study groups (healthy, caries and periodontitis) following an assessment by dental specialists. The patients' status assessment and the sample collection were performed in a previous clinical study [22]. The results from the OralDisk were compared with a lab-based extraction and qPCR reference method, as well as with the iai PadoTest (Institut für Angewandte Immunologie IAI AG, Zuchwil, Switzerland) commercial reference method. ...
... The samples used in this study were a sub-group (n = 24) of a large cohort (n = 214) of samples that had been collected at the Center for Dental Medicine, University of Zurich. These samples were intended for the microbial analysis of saliva with the aim of identifying oral infections in patients [22]. The 24 samples (seven healthy, nine caries and eight periodontitis samples) were selected in order to demonstrate the technical feasibility of the OralDisk, without the intention of generating clinical conclusions. ...
... The collection contained unstimulated whole saliva that was aliquoted and stored at −80 • C until further analysis. Details on the inclusion/exclusion criteria and on the collection methodology are available in Paqué et al. [22]. ...
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Periodontitis and dental caries are two major bacterially induced, non-communicable diseases that cause the deterioration of oral health, with implications in patients' general health. Early, precise diagnosis and personalized monitoring are essential for the efficient prevention and management of these diseases. Here, we present a disk-shaped microfluidic platform (OralDisk) compatible with chair-side use that enables analysis of non-invasively collected whole saliva samples and molecular-based detection of ten bacteria: seven periodontitis-associated (Aggregatibacter actinomycetemcomitans, Campylobacter rectus, Fusobacterium nucleatum, Prevotella intermedia, Porphyromonas gingivalis, Tannerella forsythia, Treponema denticola) and three caries-associated (oral Lactobacilli, Streptococcus mutans, Streptococcus sobrinus). Each OralDisk test required 400 µL of homogenized whole saliva. The automated workflow included bacterial DNA extraction, purification and hydrolysis probe real-time PCR detection of the target pathogens. All reagents were pre-stored within the disk and sample-to-answer processing took < 3 h using a compact, customized processing device. A technical feasibility study (25 OralDisks) was conducted using samples from healthy, periodontitis and caries patients. The comparison of the OralDisk with a lab-based reference method revealed a ~90% agreement amongst targets detected as positive and negative. This shows the OralDisk's potential and suitability for inclusion in larger prospective implementation studies in dental care settings.
... To collect biomarkers non-invasively in the oral cavity, saliva is a central favourite medium to collect and study, due to its extreme ease at sampling and plethora of parameters to analyse, including high prediction microbiological and immunological biomarkers for dental caries and periodontal disease (Paqué et al. 2020, ...
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The authors of this narrative review aimed to address various experimental methods and make recommendations for how research should move forward in the context of studying biomarkers in clinical Endodontic research. The approach adopted is exemplified using two prominent clinical problems, namely a) the "reversible" versus "irreversible" pulpitis conundrum and b) persistent idiopathic dentoalveolar pain (PIDAP). Pulpitis under deep caries or dentinal cracks is understood from a histological perspective, but clinical assessment tools to indicate irreversibly inflamed aspects of the dental pulp are elusive. PIDAP, on the other hand, is a diagnosis of exclusion; its pathophysiology is complex and not understood sufficiently to avoid unnecessary dental treatments. This review addresses how diagnostic biomarkers could further our understanding of those and other clinical problems, and how issues can be tackled from a methodological point of view. Hence, different methodological approaches to identify suitable diagnostic biomarker(s) or use known biomarkers are presented. The importance of asking a relevant research question, collecting the most suitable fluid, and using the ideal collection vehicle for the research question under investigation is discussed based on the defined clinical problems.
... Samples will be frozen at -80°C until further analysis. A. actinomycetemcomitans, P. gingivalis, T. forsythia, and T. denticola will be quantified by quantitative PCR [28]. ...
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Background Periodontal disease is a chronic state of inflammation that can destroy the supporting tissues around the teeth, leading to the resorption of alveolar bone. The initial strategy for treating periodontal disease is non-surgical sanative therapy (ST). Periodontal disease can also induce dysbiosis in the gut microbiota and contribute to low-grade systemic inflammation. Prebiotic fibers such as inulin can selectively alter the intestinal microbiota and support homeostasis by improving gut barrier functions and preventing inflammation. Providing an inulin supplement prior to and post-ST may influence periodontal health while providing insight into the complex relationship between periodontal disease and the gut microbiota. The primary objective is to determine if inulin is more effective than the placebo at improving clinical periodontal outcomes including probing depth (PD) and bleeding on probing (BOP). Secondary objectives include determining the effects of inulin supplementation pre- and post-ST on salivary markers of inflammation and periodontal-associated pathogens, as these outcomes reflect more rapid changes that can occur. Methods We will employ a single-center, randomized, double-blind, placebo-controlled study design and recruit and randomize 170 participants who are receiving ST to manage the periodontal disease to the intervention (inulin) or placebo (maltodextrin) group. A pilot study will be embedded within the randomized controlled trial using the first 48 participants to test the feasibility for the larger, powered trial. The intervention period will begin 4 weeks before ST through to their follow-up appointment at 10 weeks post-ST. Clinical outcomes of periodontal disease including the number of sites with PD ≥ 4 mm and the presence of BOP will be measured at baseline and post-ST. Salivary markers of inflammation, periodontal-associated pathogens, body mass index, and diet will be measured at baseline, pre-ST (after 4 weeks of intervention), and post-ST (after 14 weeks of intervention). Discussion We expect that inulin will enhance the positive effect of ST on the management of periodontal disease. The results of the study will provide guidance regarding the use of prebiotics prior to and as a supportive adjunct to ST for periodontal health. Trial registration ClinicalTrials.gov NCT04670133 . Registered on 17 December 2020.
... Several nucleic acid amplification technologies (NAATs) have been developed, some of them isothermal [14], while realtime PCR is still a powerful and well-established one that enables the simultaneous detection of multiple genes per reaction (including controls). POC-and lab-on-a-diskcompatible TaqMan probe duplex assays have been developed, with lyophilized amplification reagents for the scope of oral bacteria detection [15]. Quadruplex TaqMan probe assays have also been used for the detection or External Quality Assessment respiratory panels [16]. ...
... Samples will be frozen at -80°C until further analysis. A. actinomycetemcomitans, P. gingivalis, T. forsythia and T. denticola will be quanti ed by quantitative PCR [29]. ...
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Background: Periodontal disease is a chronic state of inflammation that can destroy the supporting tissues around the teeth, leading to the resorption of alveolar bone. The initial strategy for treating periodontal disease is non-surgical sanative therapy (ST). Periodontal disease can also induce dysbiosis in the gut microbiota and contribute to low grade systemic inflammation. Prebiotic fibres such as inulin can selectively alter the intestinal microbiota and support homeostasis by improving gut barrier functions and preventing inflammation. Providing an inulin supplement prior to and post-ST may influence periodontal health while providing insight into the complex relationship between periodontal disease and the gut microbiota. The primary objective is to determine if inulin is more effective than the placebo at improving clinical periodontal outcomes including probing depth (PD) and bleeding on probing (BOP). Secondary objectives include determining the effects of inulin supplementation pre and post-ST on salivary markers of inflammation and periodontal-associated pathogens, as these outcomes reflect more rapid changes that can occur. Methods: We will employ a single-center, randomized, double-blind, placebo-controlled study design and recruit and randomize 170 participants who are receiving ST to manage periodontal disease to the intervention (inulin) or placebo (maltodextrin) group. A pilot study will be embedded within the randomized controlled trial using the first 48 participants to test feasibility for the larger, powered trial. The intervention period will begin 4 weeks before ST through to their follow-up appointment at 10 weeks post-ST. Clinical outcomes of periodontal disease including number of sites with PD ≥ 4 mm and the absence of BOP will be measured at baseline and post-ST. Salivary markers of inflammation, periodontal-associated pathogens, body mass index and diet will be measured at baseline, pre-ST (after 4 weeks of intervention) and post-ST (after 14 weeks of intervention). Discussion: We expect that inulin to enhance the positive effect of ST on the management of periodontal disease. The results of the study will provide guidance regarding the use of prebiotics prior to and as a supportive adjunct to ST for periodontal health. Trial registration ClinicalTrials.gov: NCT04670133. Registered on 17 December 2020 https://clinicaltrials.gov/ct2/show/NCT04670133
... Progressing caries lesions cause an inflammation of the pulp, which leads to pulpitis and periapical periodontitis. Some bacteria, such as Streptococci and Lactobacilli, are particularly associated with the development of these caries lesions [2,12]. Persisting biofilms around the teeth on the gingival sulcus can also induce gingivitis [13,14]. ...
... 2016-00435, date of approval: 09.01.2016). A total of 120 subjects ( Figure 1) were asked to participate in a previous study (minimum age of 18 years; mean, 35.8 years; 57 females) and donate saliva for a microbial analysis and identification of diseases with a qPCR (quantitative polymerase chain reaction) assay [12]. Out of this cohort, stored (−80 • C, not longer than six months) saliva was used from healthy subjects (n = 29; mean age, 31.8; ...
... As described in a previous study [12], a strict saliva donation protocol was followed. Briefly, the participants were asked not to eat, drink sugary drinks, or perform any oral hygiene measures the night before the saliva donation. ...
Article
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This study investigated the potential of salivary bacterial and protein markers for evaluating the disease status in healthy individuals or patients with gingivitis or caries. Saliva samples from caries- and gingivitis-free individuals (n = 18), patients with gingivitis (n = 17), or patients with deep caries lesions (n = 38) were collected and analyzed for 44 candidate biomarkers (cytokines, chemokines, growth factors, matrix metalloproteinases, a metallopeptidase inhibitor, proteolytic enzymes, and selected oral bacteria). The resulting data were subjected to principal component analysis and used as a training set for random forest (RF) modeling. This computational analysis revealed four biomarkers (IL-4, IL-13, IL-2-RA, and eotaxin/CCL11) to be of high importance for the correct depiction of caries in 37 of 38 patients. The RF model was then used to classify 10 subjects (five caries-/gingivitis-free and five with caries), who were followed over a period of six months. The results were compared to the clinical assessments of dental specialists, revealing a high correlation between the RF prediction and the clinical classification. Due to the superior sensitivity of the RF model, there was a divergence in the prediction of two caries and four caries-/gingivitis-free subjects. These findings suggest IL-4, IL-13, IL-2-RA, and eotaxin/CCL11 as potential salivary biomarkers for identifying noninvasive caries. Furthermore, we suggest a potential association between JAK/STAT signaling and dental caries onset and progression.
... Thus, saliva is a very useful tool for monitoring not only oral, but also systemic health [15]. In this respect, several molecular approaches, such as PCR for RNA/DNA or ELISAs for proteins, have been proposed for detecting periodontal biomarker molecules in saliva using advanced laboratorial techniques [14,16]. However, subject to further study, simpler and more easily applicable salivary techniques may provide the pathophysiological parameters necessary for a diagnosis of periodontitis and obviate the need for blood samples or histological sections, which are traditionally required. ...
Article
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Periodontitis is a common immune-inflammatory oral disease. Early detection plays an important role in its prevention and progression. Saliva is a reliable medium that mirrors periodontal health and is easily obtainable for identifying periodontal biomarkers in point-of-care diagnostics. The aim of this study is to evaluate the effectiveness of diagnostic salivary tests to determine periodontal status. Whole saliva (stimulated/unstimulated) from twenty healthy and twenty stage III grade B generalized periodontitis patients was tested for lactoferrin, alkaline phosphatase, calcium, density, osmolarity, pH, phosphate, buffer capacity, salivary flow rate and dynamic viscosity. A semi-quantitative urinary strip test was used to evaluate markers of inflammation in saliva (erythrocytes, leukocytes, urobilinogen, nitrite, glucose, bilirubin, and ketones), clinical periodontal parameters and pathogenic bacteria. Concentrations of lactoferrin, hemoglobin, and leukocytes were found to be significantly higher in the stimulated and unstimulated saliva in periodontitis patients compared to healthy patients, whereas alkaline phosphatase levels were higher in unstimulated saliva of periodontitis patients (p < 0.05). Periodontal biomarker analysis using test strips may be considered rapid and easy tool for distinguishing between periodontitis and healthy patients. The increase in lactoferrin, hemoglobin, and leucocytes—determined by strip tests—may provide a non-invasive method of periodontal diagnosis.
... Paque et al. recently reported that the strongest differences in eight bacterial targets (C. rectus, T. forsythia, P. gingivalis, S. mutans, F. nucleatum, T. denticola, P. intermedia, and oral Lactobacilli) were found between healthy controls and periodontitis patients by their newly developed qPCR analysis of saliva (Paque et al., 2020). ...
Article
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Periodontitis is a widespread chronic inflammatory disease caused by interactions between periodontal bacteria and homeostasis in the host. We aimed to investigate the performance and reliability of machine learning models in predicting the severity of chronic periodontitis. Mouthwash samples from 692 subjects (144 healthy controls and 548 generalized chronic periodontitis patients) were collected, the genomic DNA was isolated, and the copy numbers of nine pathogens were measured using multiplex qPCR. The nine pathogens are as follows: Porphyromonas gingivalis (Pg), Tannerella forsythia (Tf), Treponema denticola (Td), Prevotella intermedia (Pi), Fusobacterium nucleatum (Fn), Campylobacter rectus (Cr), Aggregatibacter actinomycetemcomitans (Aa), Peptostreptococcus anaerobius (Pa), and Eikenella corrodens (Ec). By adding the species one by one in order of high accuracy to find the optimal combination of input features, we developed an algorithm that predicts the severity of periodontitis using four machine learning techniques. The accuracy was the highest when the models classified "healthy" and "moderate or severe" periodontitis (H vs. M-S, average accuracy of four models: 0.93, AUC = 0.96, sensitivity of 0.96, specificity of 0.81, and diagnostic odds ratio = 112.75). One or two red complex pathogens were used in three models to distinguish slight chronic periodontitis patients from healthy controls (average accuracy of 0.78, AUC = 0.82, sensitivity of 0.71, and specificity of 0.84, diagnostic odds ratio = 12.85). Although the overall accuracy was slightly reduced, the models showed reliability in predicting the severity of chronic periodontitis from 45 newly obtained samples. Our results suggest that a well-designed combination of salivary bacteria can be used as a biomarker for classifying between a periodontally healthy group and a chronic periodontitis group.
... The detection specificity to the selected resistance genes derives from the corresponding primers and potential probes. TaqMan or SYBR Green quantitative PCR assays are suitable, as they allow the monitoring of several genes of interest simultaneously [32,33]. The development of these assays is typically done in a buffer matrix, in order to avoid inhibitors for the assessment of the assay limit of detection. ...
Article
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Antibiotic resistance poses a global threat, which is being acknowledged at several levels, including research, clinical implementation, regulation, as well as by the World Health Organization. In the field of oral health, however, the issue of antibiotic resistances, as well as of accurate diagnosis, is underrepresented. Oral diseases in general were ranked third in terms of expenditures among the EU-28 member states in 2015. Yet, the diagnosis and patient management of oral infections, in particular, still depend primarily on empiric means. On the contrary, on the global scale, the field of medical infections has more readily adopted the integration of molecular-based systems in the diagnostic, patient management, and antibiotic stewardship workflows. In this perspective review, we emphasize the clinical significance of supporting in the future antibiotic resistance screening in dental practice with novel integrated and point-of-care operating tools that can greatly support the rapid, accurate, and efficient administration of oral antibiotics.