Immunoassay for H. Pylori in fecal specimens

Abstract

A process for the determination of H. Pylori in a fecal specimen comprising (a) dispersing a fecal specimen suspected of carrying H. pylori in a sample diluent; (b) contacting the fecal specimen in the diluent with a first polyclonal antibody for H. pylori antigen to form a complex of the antibody and the antigen; (c) separating said specimen and said complex; (d) exposing the complex to a second polyclonal antibody for said antigen and a portion of the antibody reacting with said complex, one of said first and second antibody being bound to a solid carrier and the other being labeled with a detecting agent; and (e) determining the amount of the labeled antibody and in turn determining the presence of H. pylori antigen in said fecal specimen.

What is claimed is:  
     
       1. A process for determination of H. pylori in a fecal specimen which comprises: 
       (a) dispersing a fecal specimen suspected of carrying H. pylori in a sample diluent;  
       (b) contacting the fecal specimen in the diluent with a first polyclonal antibody for H. pylori antigen to form a complex of the antibody and the antigen;  
       (c) separating said specimen and said complex;  
       (d) exposing the complex to a second polyclonal antibody for said antigen and a portion of the antibody reacting with said complex, one of said first and second antibody being bound to a solid carrier and the other being labelled with a detection agent; and  
       (e) determining the amount of the labelled antibody and in turn determining the presence of H. pylori antigen in said fecal specimen.  
     
     
       2. The process of  claim 1  wherein the first antibody is bound to a solid carrier and the second antibody is labelled with a detection agent. 
     
     
       3. The process of  claim 1  wherein the first antibody is labelled with a detection agent and the second is bound to a solid carrier. 
     
     
       4. The process  claim 1  wherein the sample diluent is a protein based diluent. 
     
     
       5. The process of  claim 1  wherein said polyclonal antibody is obtained by sensitizing an antibody-producing mammal with H. pylori cells. 
     
     
       6. The process of  claim 4  wherein the sample diluent contains a protein selected from the group consisting of fecal bovine serum, normal goat serum, guinea pig serum, horse serum, casein, albumin, gelatin, and bovine serum albumin. 
     
     
       7. The process of  claim 1  wherein after exposing the complex to the second antibody, the complex is washed with a buffer that reduces cross-reactivity or otherwise improves the specificity of the assay. 
     
     
       8. The process of  claim 5  wherein the cells are cells from a plurality of H. pylori strains. 
     
     
       9. The process of  claim 3  wherein said detection agent is selected from the group consisting of alkaline phosphatase and beta galactosidase horseradish peroxidase. 
     
     
       10. The process of  claim 7  wherein said wash is phosphate buffered saline. 
     
     
       11. The process of  claim 5  wherein said cells are cells from ATCC strain 43504. 
     
     
       12. A process for the determination of H. pylori in a fecal specimen which comprises: 
       (a) dispersing a fecal specimen suspected of carrying H. pylori in a sample diluent;  
       (b) contacting the fecal specimen in the diluent with a first polyclonal antibody for H. pylori antigen bound to a solid carrier and a second labelled polyclonal antibody for H. pylori to form a complex of the antibodies and the antigen;  
       (c) separating said specimen and said complex;  
       (d) determining the amount of the labelled antibody and in turn determining the presence of H. pylori antigen in said fecal specimen.  
     
     
       13. A process for the determination of H. pylori in a fecal specimen which comprises: 
       (a) dispersing a fecal specimen suspected of carrying H. pylori in a sample diluent;  
       (b) contacting the fecal specimen in the diluent with a first polyclonal antibody for H. pylori antigen produced by a first antibody-producing species and bound to a solid carrier to form a complex of the antibody and the antigen;  
       (c) separating said specimen and said complex;  
       (d) contacting the antibody-antigen complex formed in step (b) with a primary polyclonal antibody for H. pylori antigen obtained from a second antibody-producing species to produce a antibody-antigen-antibody complex;  
       (e) removing the primary antibody not present in the complex from step (d);  
       (f) contacting the antibody-antigen-antibody complex formed in step (d) with a secondary antibody, said secondary antibody being an antibody for the second antibody-producing species, whereby said secondary antibody forms a complex with said antibody-antigen-antibody complex; and  
       (g) determining the presence of H. pylori antigen in said fecal specimen.  
     
     
       14. A process for the determination of H. pylori in a fecal specimen which comprises: 
       ( a )  dispersing a fecal specimen suspected of carrying H. pylori in a sample diluent;    
       ( b )  contacting the fecal specimen in the diluent with a first polyclonal antibody for H. pylori antigen to form a complex of the first polyclonal antibody and the antigen;    
       ( c )  separating said specimen and said complex;    
       ( d )  exposing the complex to a second polyclonal antibody for said antigen and a portion of the second polyclonal antibody reacting with said complex, one of said first and second polyclonal antibody being bound to a solid carrier and the other being labeled with a detection agent; and    
       ( e )  determining the presence in said complex of the antibody labeled with the detection agent and in turn determining the presence of H. pylori antigen in said fecal specimen.   
     
     
       15. The process of  claim 14  wherein the first antibody is bound to a solid carrier and the second antibody is labeled with a detection agent. 
     
     
       16. The process of  claim 14  wherein the first antibody is labeled with a detection agent and the second is bound to a solid carrier. 
     
     
       17. The process of  claim 14  wherein the sample diluent is a protein based diluent. 
     
     
       18. The process of  claim 14  wherein said polyclonal anti- body is obtained by sensitizing an antibody - producing mammal with H. pylori cells.   
     
     
       19. The process of  claim 17  wherein the sample diluent contains a protein selected from the group consisting of fecal bovine serum, normal goat serum, guinea pig serum, horse serum, casein, albumin, gelatin, and bovine serum albumin. 
     
     
       20. The process of  claim 14  wherein after exposing the complex to the second antibody, the complex is washed with a buffer that reduces cross- reactivity or otherwise improves the specificity of the assay.   
     
     
       21. The process of  claim 18  wherein the cells are cells from a plurality of H. pylori strains. 
     
     
       22. The process of  claim 17  wherein said detection agent is selected from the group consisting of alkaline phosphatase and beta galactosidase horseradish peroxidase. 
     
     
       23. The process of  claim 20  wherein said wash is phosphate buffered saline. 
     
     
       24. The process of  claim 18  wherein said cells are cells from ATCC strain 43504. 
     
     
       25. A process for the determination of H. pylori in a fecal specimen which comprises: 
       ( a )  dispersing a fecal specimen suspected of carrying H. pylori in a sample diluent;    
       ( b )  contacting the fecal specimen in the diluent with a first polyclonal antibody for H. pylori antigen bound to a solid carrier and a second labeled polyclonal antibody for H. pylori antigen to form a complex of the first and second antibodies and the antigen;    
       ( c )  separating said specimen and said complex;    
       ( d )  determining the presence in said complex of the labeled antibody and in turn determining the presence of H. pylori antigen in said fecal specimen.