Claims for Patent: 6,599,691

Summary for Patent: 6,599,691

Title:	Rapid immunoassay to detect infection with Mycobacterium tuberculosis
Abstract:	A rapid, non-invasive, semi-quantitative immunoassay of saliva has been developed to aid in the diagnosis of diseases, e.g., using saliva to detect subjects actively or previously infected with Mycobacterium tuberculosis, a causative organism of tuberculosis. The semi-quantitative assay comprises spotting disease-related antigens on the surface of a solid substrate; contacting the solid substrate with a saliva sample which, in positive subjects, contains primary antibodies to the disease-related antigens; contacting the primary antibodies with a label capable of being detected; and detecting and reading the label whereby exposure to the antigens is determined. The device for conducting these assays is a frame or support which holds a solid substrate capable of immobilizing the antigens of interest while permitting drainage of other materials or fluids away from the immobilized antigens. A less rapid, quantitative assay has also been developed by adapting the rapid, semi-quantitative assay to an enzyme linked immunosorbant assay thereby providing a quantitative assay capable of assessing multiple saliva samples simultaneously.
Inventor(s):	Ralls; Stephen Alden (McLean, VA), Simonson; Lloyd Grant (Spring Grove, IL)
Assignee:	The United States of America as represented by the Secretary of the Navy (Washington, DC)
Application Number:	09/044,214
Patent Claims:	1. An immunodiagnostic assay kit for detecting semi-quantitatively in about five (5) minutes or quantitatively in about six (6) to eight (8) hours antibodies in saliva specific to a disease comprising: antigens specific to a disease to be identified immobilized on a solid flow-through substrate by spotting or nonspecific contact; a blocking agent for application over the antigen on said solid substrate capable of reducing nonspecific binding; a filter to remove particulate matter from a saliva sample suspected of containing primary antibodies specific to said antigens; secondary antibodies specific to said primary antibodies available for use; and a label or indicator capable of attaching directly to the primary antibodies or indirectly to said secondary antibodies producing a detectable signal. 2. The kit of claim 1 wherein the disease is any disease, condition or disorder having a detectable and specific antibody that is present or remains in saliva said antibody responding to a disease related antigen. 3. The kit of claim 2 wherein the disease is tuberculosis. 4. The kit of claim 3 wherein the microorganisms having antigens specific to a disease are selected from the group consisting of mycobacteria and derivatives, Mycobacterium tuberculosis, Mycbacterium bovis, tuberculin purified protein derivative, the Bacillus of Calmette and Guerin and lipoarabinomannan of Mycobacterium tuberculosis. 5. The kit of claim 1 wherein the label or indicator is selected from the group consisting of colloidal gold; colloidal gold coupled to a protein; an enzyme; a fluorescent marker; a radionuclide; and latex particles. 6. The kit of claim 5 wherein the label is alkaline phosphatase. 7. The kit of claim 5 wherein the protein coupled to the colloidal gold is selected from the group consisting of protein-A and protein-G. 8. An immunodiagnostic assay method for detecting antibodies in saliva specific to a disease comprising: contacting and immobilizing antigens specific to a disease with a solid flow-through substrate to form a spot or as nonspecific contact; blocking said solid substrate to reduce nonspecific binding; gathering a saliva sample suspected of containing primary antibodies to the antigens specific to a disease; separating particulate matter from said sample with a separating device selected from the group consisting of filters and centrifuges to form a salivary sample filtrate or supernatant; spotting the saliva filtrate or supernatant on to the immobilized antigen on the solid substrate; contacting the immobilized antigen and sample on the solid substrate with a label capable of directly attaching to the primary antibodies or indirectly attaching to secondary antibodies specific to the primary antibodies; and detecting and reading the intensity of the label in less than 5 minutes whereby the presence and concentration of primary antibodies in the sample are determined. 9. The method of claim 8 wherein the disease is any disease, condition or disorder having a specific, detectable host antibody response. 10. The method of claim 9 wherein the disease is tuberculosis. 11. The method of claim 10 wherein the microorganisms containing antigens specific to a disease are selected from the group consisting of mycobacteria and derivatives, Mycobacterium tuberculosis, Mycobacterium bovis, tuberculin purified protein derivative, the Bacillus of Calmette and Guerin and lipoarabinomannan of Mycobacterium tuberculosis. 12. The method of claim 8 wherein the label or indicator is selected from the group consisting of colloidal gold; colloidal gold coupled to a protein; an enzyme; a fluorescent marker; a radionuclide; and latex particles. 13. The method of claim 12 wherein the label is alkaline phosphatase. 14. The method of claim 12 wherein the protein coupled to the colloidal gold is selected from the group consisting of protein-A and protein-G.

Details for Patent 6,599,691

Applicant	Tradename	Biologic Ingredient	Dosage Form	BLA	Approval Date	Patent No.	Expiredate
Par Pharmaceutical Companies, Inc.	APLISOL	tuberculin, purified protein derivative	Injection	103782	04/20/1998		2015-11-27
Sanofi Pasteur Limited	TUBERSOL	tuberculin, purified protein derivative	Injection	103941	02/24/2000		2015-11-27