Methods and apparatus for detection of gluten sensitivity, and its differentiation from celiac disease

Abstract

Antibodies are used as biomarkers to assist in distinguishing gluten immune reactivity and sensitivity, silent celiac disease, Crohn's disease and other gut-related pathologies from classical celiac disease. In one class of embodiments, sera, saliva or other samples from a human or other animal are tested for antibodies to (a) a wheat antigen; (b) a gliadin antigen; and (c) one or more of a wheat germ agglutinin, a gluteomorphin, a glutenin, a deamidated glutenin, a prodynorphin, and a dynorphin. Test results are considered particularly interesting where the wheat antigen and the gliadin antigen are both selected from the group consisting of native and deamidated forms of α-gliadin 33-mer, α-gliadin-17-mer, γ-gliadin-15-mer, ω-gliadin-17-mer, and glutenin 21-mer. Test plates and kits can advantageously test for antibodies to at least three, five, seven or all of mixed wheat antigens, α-gliadin, γ-gliadin, ω-gliadin, glutenin, α-glutenin, wheat germ agglutinin, gluteomorphin, prodynorphins, transglutaminase-2, transglutaminase-3, transglutaminase-6, and gliadin-bound transglutaminase.

1 - 9 . (canceled) 
     
     
         10 . A device for characterizing wheat sensitivity in a subject, comprising:
 a first test region comprising an antigenic α-gliadin peptide;   a second test region comprising an antigenic deamidated α-gliadin peptide;   a third test region comprising an antigenic glutenin peptide;   a fourth test region comprising an antigenic deamidated glutenin peptide; and   a fifth test region comprising an antigenic complex comprising gliadin and transglutaminase.   
     
     
         11 . The device of  claim 10 , wherein the antigenic α-gliadin peptide comprises 33 or fewer amino acids, and the deamidated α-gliadin peptide comprises 33 or fewer amino acids. 
     
     
         12 . The device of  claim 11 , wherein the antigenic α-gliadin peptide is selected from the group consisting of α-gliadin 33-mer of SEQ ID NO. 1 and α-gliadin 17-mer of SEQ ID NO.6, and the deamidated α-gliadin peptide is selected from the group consisting of deamidated α-gliadin 33-mer of SEQ ID NO. 2 and deamidated α-gliadin 17-mer of SEQ ID NO.7. 
     
     
         13 . The device of  claim 10 , wherein the antigenic glutenin peptide comprises 21 or fewer amino acids, and the antigenic deamidated glutenin peptide comprises 21 or fewer amino acids. 
     
     
         14 . The device of  claim 13 , wherein the antigenic glutenin peptide comprises glutenin 21-mer of SEQ ID NO. 15, and the antigenic deamidated glutenin peptide comprises deamidated glutenin 21-mer of SEQ ID NO. 16. 
     
     
         15 . The device of  claim 10 , wherein the antigenic complex is selected from the group consisting of gliadin-transglutaminase complex A of SEQ ID NO. 19 and gliadin-transglutaminase complex B of SEQ ID NO.20. 
     
     
         16 . The device of  claim 10 , further comprising a sixth test region comprising at least one of an antigenic γ-gliadin peptide and an antigenic ω-gliadin peptide. 
     
     
         17 . The device of  claim 10 , further comprising a sixth test region comprising an antigenic gluteomorphin peptide. 
     
     
         18 . The device of  claim 10 , further comprising a sixth test region comprising an antigenic prodynorphin peptide. 
     
     
         19 . The device of  claim 10 , further comprising a sixth test region comprising an antigenic wheat germ agglutinin peptide. 
     
     
         20 . A method of characterizing wheat sensitivity in a subject, comprising:
 identifying the presence of a first antibody of the subject in a sample from the subject, wherein the first antibody forms a first complex with an antigenic α-gliadin peptide;   identifying the presence of a second antibody of the subject in the sample, wherein the second antibody forms a second complex with an antigenic deamidated antigenic α-gliadin peptide;   identifying the presence of third antibody of the subject in the sample, wherein the third antibody forms a third complex with an antigenic glutenin peptide;   identifying the presence of a fourth antibody of the subject in the sample, wherein the fourth antibody forms a fourth complex with an antigenic deamidated glutenin peptide; and   identifying the presence of a fifth antibody of the subject in the sample, wherein the fifth antibody forms a fifth complex with an antigenic complex comprising gliadin and transglutaminase.   
     
     
         21 . The method of  claim 20 , wherein the antigenic α-gliadin peptide comprises 33 or fewer amino acids, and the deamidated α-gliadin peptide comprises 33 or fewer amino acids. 
     
     
         22 . The method of  claim 21 , wherein the antigenic α-gliadin peptide is selected from the group consisting of α-gliadin 33-mer of SEQ ID NO. 1 and α-gliadin 17-mer of SEQ ID NO. 6, and the deamidated α-gliadin peptide is selected from the group consisting of deamidated α-gliadin 33-mer of SEQ ID NO. 2 and deamidated α-gliadin 17-mer of SEQ ID NO. 7. 
     
     
         23 . The method of  claim 20 , wherein the antigenic glutenin peptide comprises 21 or fewer amino acids, and the antigenic deamidated glutenin peptide comprises 21 or fewer amino acids. 
     
     
         24 . The method of  claim 23 , wherein the antigenic glutenin peptide comprises glutenin 21-mer of SEQ ID NO. 15, and the antigenic deamidated glutenin peptide comprises deamidated glutenin 21-mer of SEQ ID NO. 16. 
     
     
         25 . The method of  claim 20 , wherein the antigenic complex is selected from the group consisting of gliadin-transglutaminase complex A of SEQ ID NO. 19 and gliadin-transglutaminase complex B of SEQ ID NO.20. 
     
     
         26 . The method of  claim 20 , further comprising a sixth test region comprising at least one of an antigenic γ-gliadin peptide and an antigenic ω-gliadin peptide. 
     
     
         27 . The method of  claim 20 , further comprising a sixth test region comprising an antigenic gluteomorphin peptide. 
     
     
         28 . The method of  claim 20 , further comprising a sixth test region comprising an antigenic prodynorphin peptide. 
     
     
         29 . The method of  claim 20 , further comprising a sixth test region comprising an antigenic wheat germ agglutinin peptide.