Detection of oral microbial virulence factors

Abstract

Methods for detecting and quantifying toxins present in the oral cavity. The methods may include providing a biological sample, providing reporter cells expressing one or more Toll like receptors, exposing the cells to the biological sample, measuring the EC50 value of the lipopolysaccharide on activation of a Toll like receptor, quantification of the lipopolysaccharide in the biological sample.

What is claimed is: 
     
       1. A method of determining bacterial virulence in an oral cavity comprising:
 a. providing a reporter cell expressing at least one toll-like receptor; 
 b. providing a sample of oral matter; 
 c. combining the sample of oral matter and the reporter cell; 
 d. measuring the toll-like receptor activation; 
 e. providing another sample of oral matter from an individual who used an oral care composition prior to providing another sample of oral matter; 
 f. combining the another sample of oral matter and the reporter cell; 
 g. measuring the toll-like receptor response; 
 h. comparing the toll-like receptor response from the sample of oral matter and another sample of oral matter. 
 
     
     
       2. The method of  claim 1 , wherein the toll-like receptor is at least one of TLR1 to TLR9. 
     
     
       3. The method of  claim 1 , wherein the reporter cell expresses at least of one TLR2 or TLR4. 
     
     
       4. The method of  claim 1 , wherein the toll-like receptor activates a reporter gene. 
     
     
       5. The method of  claim 4 , wherein the reporter gene is a secreted embryonic alkaline phosphatase (SEAP) reporter gene. 
     
     
       6. The method of  claim 1 , wherein the sample of oral matter is at least one of gum-line plaque, subgingival plaque, supragingival plaque, interstitial plaque, gingival crevicular fluid (GCF), gingival biopsy, saliva, or tongue swab. 
     
     
       7. The method of  claim 1 , wherein the sample of oral matter comprises virulence factor that is at least one of Gram positive cell wall components, Gram negative cell wall components, bacterial DNA, flagellin, peptidoglycan, bacterial metabolites, fimbriae; outer membrane vesicles bacterial proteins, or bacterial enzymes. 
     
     
       8. The method of  claim 7 , wherein the sample of oral matter comprises virulence factor that is at least one of lipopolysaccharide or lipoteichoic acid. 
     
     
       9. The method of  claim 1 , where the sample of oral matter and the another sample of oral matter are from the same individual.