Intended use

Enzyme immunoassay for the detection of autoantibodies IgA against modified gliadin peptide (MGP).

Background

Celiac disease (CD) is caused by a hypersensitive reaction of genetically predisposed individuals to ingested gluten. Gluten is a set of proteins present in many kinds of cereal grain, e.g. wheat, barley and rye. CD affects the upper small intestine and results in more or less complete atrophy of the villi of the mucous membrane which can lead to malabsorption problems, e.g. chronic vitamin deficiency. The symptoms are variable or sometimes even absent.

Elevated levels of gliadin-specific antibodies occur in the sera of celiac patients. Gliadin is a component of gluten and constitutes a predominant antigen. It is fragmented in the small intestine and the resulting peptides are deamidated by the enzyme tissue transglutaminase (tTG) which itself has been identified as the major CD autoantigen.

Recently, it was shown that certain deamidated gliadin peptides are powerful immunogens and that antibodies directed at these peptides exhibit a better diagnostic accuracy for CD, as compared to antibodies directed at crude gliadin.

Technical information

The EULISA Modified Gliadin Peptide IgA is intended for the quantitative or qualitative determination of IgA antibodies in human serum, directed against a modified (deamidated) gliadin peptide (MGP). The immobilised antigen is a highly purified, synthetic peptide derivative. The test is fast (incubation time 30-30-30 minutes) and flexible (break apart wells, ready-to-use reagents). Six calibrators allow quantitative measurements; a negative and a positive control are used to check assay performance.

Kit components and storage of reagents

Contents of the kit

1. One microwell plate, coated with MGP and hermetically packed in a foil laminate pouch together with a desiccant bag.

2. Sample buffer, 100 mL, ready-to-use, orange coloured.

3. Wash buffer, 100 mL, 10 x-concentrate, blue coloured.

4. 6 calibrators, 2.0 mL each, 0, 3.0, 8.0, 18, 45 and 100 U MGP antibodies IgA/mL, ready-to-use, gradually blue coloured.

5. Negative and positive control, 2.0 mL each, ready-to-use, green and red coloured, respectively.

6. Anti-human IgA HRP conjugate, 14 mL, ready-to-use, red coloured.

7. Substrate solution, 14 mL, ready-to-use, colourless.

8. Stop solution (0.5 M H2SO4), 14 mL, colourless, ready-to-use.

9. Directions for use

10. Lot-specific certificate of analysis

All kit components are stored at 2 - 8°C.