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US-12618854-B2 - Diagnostics of mild or adversed periodontitis

US12618854B2US 12618854 B2US12618854 B2US 12618854B2US-12618854-B2

Abstract

Disclosed is an in vitro method for assessing whether a human patient suffering from periodontitis has mild periodontitis or advanced periodontitis. The method is based on the insight to determine a selection of two biomarker proteins. Accordingly, in a sample of saliva a patient suffering from periodontitis, the concentrations are measured of the proteins Pyruvate Kinase (PK) and at least one of Matrix metalloproteinase-9 (MMP9), S100 calcium-binding protein A8 (S100A8), and Hemoglobin subunit beta (Hb-beta); or of the proteins Matrix metalloproteinase-9 (MMP9) and at least one of S 100 calcium-binding protein A8 (S100A8) and S100 calcium-binding protein A9 (S100A9). Based on the concentrations as measured, a value is determined reflecting the joint concentrations for said proteins. This value is compared with a threshold value reflecting in the same manner the joint concentrations associated with advanced periodontitis. The comparison allows assessing whether the testing value is indicative of the presence of advanced periodontitis or of mild periodontitis in said patient. Thereby, typically, a testing value reflecting a joint concentration below the joint concentration reflected by the threshold value is indicative for mild periodontitis in said patient, and a testing value reflecting a joint concentration at or above the joint concentration reflected by the threshold value, is indicative for advanced periodontitis in said patient.

Inventors

  • Gerben Kooijman
  • AMIR HUSSEIN RMAILE
  • Carl Glasse
  • Marinus Karel Johannes De Jager
  • IAIN LESLIE CAMPBELL CHAPPLE
  • MELISSA MACKAY GRANT
  • PHILIP PRESHAW
  • John Taylor
  • Michael Alex Van Hartskamp

Assignees

  • KONINKLIJKE PHILIPS N.V.

Dates

Publication Date
20260505
Application Date
20190410

Claims (20)

  1. 1 . A method for treating a human patient who has mild periodontitis or advanced periodontitis, wherein the method comprises: detecting, in a sample of saliva from said human patient, the concentrations of the proteins: Pyruvate Kinase (PK) and Matrix metalloproteinase-9 (MMP9); determining a testing value reflecting the joint concentrations determined for said proteins; and comparing said testing value with a threshold value reflecting in the same manner the joint concentrations associated with advanced periodontitis, so as to assess that the testing value is indicative for mild periodontitis or for advanced periodontitis in said patient; and administering, to the human patient for which the testing value is indicative for mild periodontitis or for advanced periodontitis, an anti-microbial therapeutic agent to the gum tissue of the human patient.
  2. 2 . The method of claim 1 , wherein the human patient is known to have periodontitis.
  3. 3 . The method of claim 1 , wherein an age of the subject is determined and the testing value reflects the joint concentrations determined for said proteins, in combination with the age of the subject.
  4. 4 . The method of claim 3 , wherein the proteins further comprise S100 calcium-binding protein A8 (S100A8).
  5. 5 . The method of claim 3 , wherein the proteins further comprise Hemoglobin subunit beta (Hb-beta).
  6. 6 . The method of claim 1 , wherein the threshold value is based on the concentrations determined for the proteins in one or more reference samples each sample associated with the presence of advanced periodontitis.
  7. 7 . The method of claim 1 , wherein the threshold value is based on the concentrations of the proteins in a set of samples, including samples from subjects that have mild or moderate periodontitis and samples from subjects having advanced periodontitis.
  8. 8 . The method of claim 1 , wherein the concentration values determined are arithmetically processed into a number between 0 and 1.
  9. 9 . A method of treating a human patient who has mild periodontitis or advanced periodontitis, comprising; detecting in a sample of saliva of the human patient the proteins: Pyruvate Kinase (PK) and Matrix metalloproteinase-9 (MMP9); determining a testing value reflecting joint concentrations determined for said proteins; and assessing the presence of mild periodontitis or advanced periodontitis in the patient on the basis of the concentrations of said proteins in said sample, comprising comparing said determined testing value with a threshold value reflecting in the same manner the joint concentrations associated with advanced periodontitis, so as to assess whether the testing value is indicative for mild periodontitis or for advanced periodontitis in said patient; and administering, to the human patient for which the assessment is indicative for mild periodontitis or for advanced periodontitis, an anti-microbial therapeutic agent to the gum tissue of the human patient.
  10. 10 . The method of claim 9 , wherein the human patient is known to have periodontitis.
  11. 11 . The method of claim 9 , wherein an age of the subject is determined and the testing value reflects the joint concentrations determined for said proteins, in combination with the age of the subject.
  12. 12 . The method of claim 11 , wherein the proteins further comprise S100 calcium-binding protein A8 (S100A8).
  13. 13 . The method of claim 11 , wherein the proteins further comprise Hemoglobin subunit beta (Hb-beta).
  14. 14 . The method of claim 9 , wherein the threshold value is based on the concentrations determined for the proteins in one or more reference samples each sample associated with the presence of advanced periodontitis.
  15. 15 . The method of claim 9 , wherein the threshold value is based on the concentrations of the proteins in a set of samples, including samples from subjects that have mild or moderate periodontitis and samples from subjects having advanced periodontitis.
  16. 16 . The method of claim 9 , wherein the concentration values determined are arithmetically processed into a number between 0 and 1.
  17. 17 . A method of treating periodontitis comprising: detecting the proteins: Pyruvate Kinase (PK) and Matrix metalloproteinase-9 (MMP9) in a human patient, by: (a) obtaining a saliva sample from a human patient; and (b) detecting whether the proteins are present in the sample by contacting the sample with one or more detection reagents for binding said proteins and detecting binding between each protein and the one or more detection reagents; and administering, to the human patient for which the proteins are detected in concentrations indicative for mild periodontitis or for advanced periodontitis, an anti-microbial therapeutic agent to the gum tissue of the human patient.
  18. 18 . The method of claim 17 , wherein the human patient is known to have periodontitis.
  19. 19 . The method of claim 17 , wherein an age of the subject is determined and the testing value reflects the joint concentrations determined for said proteins, in combination with the age of the subject.
  20. 20 . The method of claim 19 , wherein the proteins further comprise S100 calcium-binding protein A8 (S100A8).

Description

CROSS-REFERENCE TO PRIOR APPLICATIONS This application is the U.S. National Phase application under 35 U.S.C. § 371 of International Application No. PCT/EP2019/059050, filed on Apr. 10, 2019, which claims the benefit of European Patent Application No. 18166969.8, filed on Apr. 12, 2018. These applications are hereby incorporated by reference herein. FIELD OF THE INVENTION The invention is in the field of oral care, and pertains to saliva-based diagnostics of periodontal disease. Particularly, the invention pertains to a kit and method for distinguishing mild from advanced periodontitis. BACKGROUND OF THE INVENTION Gum inflammation, or gingivitis, is a non-destructive periodontal disease caused mainly by the adherence of dental bacterial biofilms, or dental plaque, to the tooth surface that triggers an inflammatory reaction in the surrounding tissue. Gingivitis is a reversible infection and inflammation of the gum tissues, and may be resolved with proper oral hygiene measures and dental professional intervention. If not detected or resolved, gingivitis usually leads to the inflammation of the tissues surrounding the tooth (i.e. periodontal tissues), a condition defined as periodontitis that causes tissue destruction and alveolar bone loss, and ultimately results in the loss of teeth. During the progression of gum disease, there are usually clinical signs and symptoms associated with it, such as the swelling of the gums, the change in color from pink to dark red, the bleeding of the gums, bad breath, and the gums becoming more tender or painful to touch. Periodontitis is a chronic multifactorial inflammatory disease caused by oral microorganisms and characterized by progressive destruction of the hard (bone) and soft (periodontal ligament) tissues, ultimately leading to tooth mobility and loss. This is to be distinguished from gingivitis which is a reversible infection and inflammation of the gum tissues. Inflammatory periodontitis is one of the most prevalent chronic human diseases and a major cause of adult tooth loss. In addition to the substantial negative impact of periodontitis on oral health, there is also mounting evidence that periodontitis has systemic consequences and that it is a risk factor for several systemic diseases, including heart diseases (e.g. atherosclerosis, stroke), diabetes, pregnancy complications, rheumatoid arthritis and respiratory infections. Early and accurate diagnosis of periodontal disease, thus, is important from both an oral and overall health perspective. Periodontal diseases are still poorly diagnosed in general dental practice, resulting in relatively low rates of therapeutic intervention and significant amounts of untreated cases. Current diagnosis relies on imprecise, subjective clinical examination of oral tissue condition (color, swelling, extent of bleeding on probing, probing pocket depth; and bone loss from oral x-rays) by dental professionals. These conventional methods are time consuming, and some of the techniques used (pocket-depth, x-ray) reflect historic events, such as past disease activity, rather than current disease activity or susceptibility to further disease. Hence, more objective, faster, accurate, easier-to-use diagnostics which preferably may also be performed by non-specialists are desirable. Thereby it is desirable to measure current disease activity, and possibly a subject's susceptibility to further periodontal disease. Saliva or oral fluids have long been advocated as a diagnostic fluid for oral and general diseases, and with the advent of miniaturized biosensors, also referred to as lab-on-a-chip, point of care diagnostics for rapid chair-side testing have gained greater scientific and clinical interest. Especially for periodontal disease detection, inflammatory biomarkers associated with tissue inflammation and breakdown may easily end up in saliva due to proximity, suggesting saliva has strong potential for periodontal disease detection. Indeed, this area thus has gained significant interest and encouraging results have been presented. For example, Kido et al. (J Periodont Res 2012; 47:488-499) identified 104 proteins in gingival crevicular fluid (GCF) samples from both healthy sites and sites of periodontitis, 64 proteins contained only in GCF from healthy sites and 63 proteins only in GCF from periodontitis sites. However, no definite test has emerged yet. Biomarkers represent biological indicators that underpin clinical manifestations, and as such are objective measures by which to diagnose clinical outcomes of periodontal disease. Ultimately, proven biomarkers could be utilized to assess risk for future disease, to identify disease at the very earliest stages, to identify response to initial therapy, and to allow implementation of preventive strategies. Previous limitations to the development of point-of-care tests for salivary biomarkers included a lack of technologies that were adaptable to chair-side applications and an inability to