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 for determining the virulence of lipopolysaccharide comprising:
 a. providing a lipopolysaccharide; 
 b. providing a reporter cell expressing at least one Toll-like receptor; 
 c. combining the reporter cell with the lipopolysaccharide; 
 d. measuring the toll-like receptor activation; 
 e. quantifying the lipopolysaccharide; and 
 wherein the Toll-like receptor activation is determined by measuring at least one of EC50 or fold of stimulation. 
 
     
     
       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 one of 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 reported gene is a secreted embryonic alkaline phosphatase (SEAP) reporter gene. 
     
     
       6. The method of  claim 1 , wherein the lipopolysaccharide is quantified using at least one of limulus amebocyte lysate assay or a fluorescence assay. 
     
     
       7. A method for determining the virulence of lipoteichoic acid comprising:
 a. providing a lipoteichoic acid; 
 b. providing a reporter cell expressing at least one Toll-like receptor; 
 c. combining the reporter cell with the lipoteichoic acid; 
 d. measuring the toll-like receptor activation; 
 e. quantifying the lipoteichoic acid; and 
 wherein the Toll-like receptor activation is determined by measuring at least one of EC50 or fold of stimulation. 
 
     
     
       8. The method of  claim 7 , wherein the Toll-like receptor is at least one of TLR1 to TLR9.