Introduction: More and more evidence has accumulated that suggests salivary sampling may provide direct analysis of oral conditions and microbial constituents, but may also be useful in the diagnosis and early detection of other chronic diseases. Although multiple methods of oral sampling currently exist, some methods are prohibitively expensive or based upon technologies not ubiquitously available at public health centers or state-funded colleges. This study provides a comparative analysis of DNA concentrations and quality from five specific oral sites derived using sterile paper points, including the gingival crevice between the upper central incisors, biofilm of the upper first molar, lingual incisor, and the dorsum of the tongue for comparison with unstimulated saliva collection. Methods: This study analyzed previously collected unstimulated saliva and paper point samples. In brief, DNA was isolated from each using TRIzol (phenol: Chloroform) extraction and DNA quantification and quality was measured using a NanoDrop spectrophotometer at 260 and 280 nm. Results: Analysis of Paper Point (PP) biofilm sampling sites from upper first molar, lower incisor, and dorsum of the tongue revealed similar average DNA concentrations, ranging between 14,342 ng/uL and 14,402 ng/uL (p=0.9851). Although variations were observed between different patients, samples from different oral sites within the same patient were strikingly similar, R=0.8355. Comparison of DNA isolated from fluids, gingival crevicular fluid (GCF) and unstimulated saliva revealed average DNA concentrations that were similar to the biofilm sampling sites (14,686 ng/uL and 13,743 ng/uL, respectively), which were not significantly different from one another (p=0.7893). DNA concentrations ranged considerably between patients (low = 4,410 ng/uL; high = 48,783 ng/uL), but were most similar with different samples (GCF, saliva) from the same patient (Pearson’s R=0.6979). In addition, DNA purity measured by A260:A280 nm absorbance did not reveal any significant difference among sampling sites (range 1.62 – 1.70; p=0.427). Discussion: Although many methods are available to provide oral sampling, simple and low-cost methods such as paper point sampling, unstimulated saliva collection and buccal swabs may represent tools that provide sufficient DNA quality and quantity for molecular screening. In addition, although heterogeneity between patient samples will always be present – samples from various oral sites within the same patient may provide roughly equivalent DNA samples for further screening and molecular analysis.

Introduction: The newly discovered cariogenic pathogen Scardovia wiggsiae has been the subject of intense scientific interest due to the role it may play in the development or progression of caries and oral disease. The primary objective of this study was to perform DNA microbial screening from five specific oral sites, including the gingival crevice between the upper central incisors, biofilm of the upper first molar and lingual incisor, as well as the dorsum of the tongue – for comparison with unstimulated saliva. These data may provide significant insights into site-specific oral locations that harbor S. wiggsiae. Methods: More than one hundred previously collected clinical samples (n=105) were identified for inclusion in this study. DNA isolates were screened using a NanoDrop spectrophotometer to determine overall DNA quantity and quality. Samples with sufficient quality and quantity were screened for the presence of S. wiggsiae using validated PCR primers. Results: More than one hundred patient samples (n=105) were identified, which were comprised of mostly female (57%) versus male (43%) and minority (71%) versus White (29%). The average DNA concentrations ranged between 13.74 and 14.69 mg/mL, with A260:A280 ratios ranging between 1.62 – 1.70. Results of molecular screening using S. wiggsiae specific primers demonstrated only a small percentage of pooled samples (7.6%) harbored this DNA, which was highly concentrated among the samples from tooth surfaces (Upper First Molar, Lingual Incisor) and saliva compared with the gingival crevice and dorsum of the tongue. Discussion: These data provide novel information regarding specific oral locations, including tooth surfaces that harbor S. wiggsiae. In addition, these sites also provide new information regarding oral sites that do not appear to harbor this organism, including the gingival crevice and dorsum of the tongue. This information may be particularly useful to oral health researchers as they strive to limit and reduce the cariogenic microbiome among high-risk populations.