Discovery, validation, and diagnostic ability of multiple protein-based biomarkers in saliva and gingival crevicular fluid to distinguish between health and periodontal diseases

J Clin Periodontol. 2022 Jul;49(7):622-632. doi: 10.1111/jcpe.13630. Epub 2022 Apr 29.


Aim: To discover and validate differential protein biomarker expression in saliva and gingival crevicular fluid (GCF) to discriminate objectively between periodontal health and plaque-induced periodontal disease states.

Materials and methods: One-hundred and ninety participants were recruited from two centres (Birmingham and Newcastle upon Tyne, UK) comprising healthy, gingivitis, periodontitis, and edentulous donors. Samples from the Birmingham cohort were analysed by quantitative mass spectrometry proteomics for biomarker discovery. Shortlisted candidate proteins were then verified by enzyme-linked immunosorbent assay in both cohorts. Leave-one-out cross validation logistic regression analysis was used to identify the best performing biomarker panels.

Results: Ninety-five proteins were identified in both GCF and saliva samples, and 15 candidate proteins were selected based upon differences discovered between the donor groups. The best performing panels to distinguish between: health or gingivitis and periodontitis contained matrix metalloproteinase-9 (MMP9), S100A8, alpha-1-acid glycoprotein (A1AGP), pyruvate kinase, and age (area under the curve [AUC] 0.970); health and gingivitis contained MMP9, S100A8, A1AGP, and pyruvate kinase, but not age (AUC 0.768); and mild to moderate and advanced periodontitis contained MMP9, S100A8, A1AGP, pyruvate kinase, and age (AUC 0.789).

Conclusions: Biomarker panels containing four proteins with and without age as a further parameter can distinguish between periodontal health and disease states.

Keywords: biomarker; gingival crevicular fluid; periodontal disease; proteomics; saliva.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Biomarkers / analysis
  • Chronic Periodontitis* / metabolism
  • Gingival Crevicular Fluid / chemistry
  • Gingivitis* / diagnosis
  • Gingivitis* / metabolism
  • Humans
  • Matrix Metalloproteinase 9 / analysis
  • Pyruvate Kinase / analysis
  • Saliva / chemistry


  • Biomarkers
  • Pyruvate Kinase
  • Matrix Metalloproteinase 9