Microbial Analysis of Saliva to Identify Oral Diseases Using a Point-of-Care Compatible qPCR Assay

Abstract

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 Lactobacilli) in saliva and their clinical status association in 214 individuals. Upon clinical examination, study subjects were grouped into healthy, caries and periodontitis and their saliva was collected. A highly specific point-of-care compatible dual color qPCR assay was developed and used to study the above-mentioned bacteria of interest in the collected saliva. Assay performance was compared to a commercially available microbial reference test. Eight out of ten taxa that were investigated during this study were strong discriminators between the periodontitis and healthy groups: C. rectus, T. forsythia, P. gingivalis, S. mutans, F. nucleatum, T. denticola, P. intermedia and oral Lactobacilli (p < 0.05). Significant differentiation between the periodontitis and caries group microbiome was only shown for S. mutans (p < 0.05). A clear distinction between oral health and disease was enabled by the analysis of quantitative qPCR data of target taxa levels in saliva.

Salivary Biomarkers for Dental Caries Detection and Personalized Monitoring

“…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; 18 females), patients with gingivitis ( n = 22; mean age, 35.4; 13 females), and patients with open caries lesions ( n = 69; mean age, 37.6; 26 females).…”

“…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 ].…”

Healthcare Challenges and Future Solutions in Dental Practice: Assessing Oral Antibiotic Resistances by Contemporary Point-Of-Care Approaches

“…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.…”

Prediction of Chronic Periodontitis Severity Using Machine Learning Models Based On Salivary Bacterial Copy Number

“…Morozumi et al evaluated the amount of bacteria in saliva and IgG titers in serum to monitor the progression of chronic periodontitis (Morozumi et al, 2016). 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).…”

“…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 ).…”