title: Microbial signatures of oral dysbiosis, periodontitis and edentulism revealed by Gene Meter methodology.
abstract: Conceptual models suggest that certain microorganisms (e.g., the "red" complex) are indicative of a specific disease state (e.g., periodontitis); however, recent studies have questioned the validity of these models. Here, the abundances of 500+ microbial species were determined in 16 patients with clinical signs of one of the following oral conditions: periodontitis, established caries, edentulism, and oral health. Our goal was to determine if the abundances of certain microorganisms reflect dysbiosis or a specific clinical condition that could be used as a 'signature' for dental research. Microbial abundances were determined by the analysis of 138,718 calibrated probes using Gene Meter methodology. Each 16S rRNA gene was targeted by an average of 194 unique probes (n=25nt). The calibration involved diluting pooled gene target samples, hybridizing each dilution to a DNA microarray, and fitting the probe intensities to adsorption models. The fit of the model to the experimental data was used to assess individual and aggregate probe behavior; good fits (R2>0.90) were retained for back-calculating microbial abundances from patient samples. The abundance of a gene was determined from the median of all calibrated individual probes or from the calibrated abundance of all aggregated probes. With the exception of genes with low abundances (<2 arbitrary units), the abundances determined by the different calibrations were highly correlated (r~1.0). Seventeen genera were classified as 'signatures of dysbiosis' because they had significantly higher abundances in patients with periodontitis and edentulism when contrasted with health. Similarly, 13 genera were classified as 'signatures of periodontitis', and 14 genera were classified as 'signatures of edentulism'. The signatures could be used, individually or in combination, to assess the clinical status of a patient (e.g., evaluating treatments such as antibiotic therapies). Comparisons of the same patient samples revealed high false negatives (45%) for next-generation-sequencing results and low false positives (7%) for Gene Meter results.
authors.affiliation: Program in Microbiology, Alabama State University, Montgomery, AL 36101, United States. Electronic address: cmooghww@gmail.com.
authors.forename: M Colby
authors.initials: MC
authors.lastname: Hunter
authors.affiliation: Department of Oral Health, University of Washington, Box 3574444, Seattle, WA, United States. Electronic address: pozhit@uw.edu.
authors.forename: Alex E
authors.initials: AE
authors.lastname: Pozhitkov
authors.affiliation: Department of Periodontics, University of Washington, Box 3574444, Seattle, WA, United States. Electronic address: panoble@washington.edu.
authors.forename: Peter A
authors.initials: PA
authors.lastname: Noble
mesh_terms: D000328:Adult; D001419:Bacteria / Q000145:classification* / Q000235:genetics* / Q000302:isolation & purification*; D001483:Base Sequence; D002138:Calibration; D015342:DNA Probes; D004269:DNA, Bacterial / Q000235:genetics; D003731:Dental Caries / Q000175:diagnosis / Q000382:microbiology; D003773:Dental Plaque / Q000382:microbiology; D064806:Dysbiosis / Q000175:diagnosis / Q000382:microbiology*; D005188:False Negative Reactions; D005189:False Positive Reactions; D005260:Female; D018390:Gene Targeting / Q000379:methods; D059014:High-Throughput Nucleotide Sequencing; D006801:Humans; D008297:Male; D008828:Microbiological Techniques / Q000379:methods*; D064307:Microbiota / Q000235:genetics; D008875:Middle Aged; D025202:Molecular Diagnostic Techniques / Q000379:methods*; D009055:Mouth / Q000382:microbiology*; D009693:Nucleic Acid Hybridization / Q000379:methods; D020411:Oligonucleotide Array Sequence Analysis / Q000379:methods / Q000592:standards; D009909:Oral Health; D010518:Periodontitis / Q000175:diagnosis / Q000382:microbiology*; D012336:RNA, Ribosomal, 16S / Q000235:genetics; D017421:Sequence Analysis; D017422:Sequence Analysis, DNA / Q000379:methods; D014861:Washington
chemical_list: D015342:DNA Probes; D004269:DNA, Bacterial; D012336:RNA, Ribosomal, 16S
keywords: 454 sequencing; Calibrated DNA microarrays; Caries; Edentulism; Gene Meter; Microbial abundance signatures; Next-generation sequencing; Periodontitis