Gluten ELISA Kit (Gliadin)

Full Name: Gluten ELISA Kit (Gliadin)
Sample Type: Food (Baby Food, Chocolate, Bakery, Beer, Meat)
Sensitivity: 0.3 ppm

BACKGROUND

Gluten is found in grains such as; wheat, barley, rye and spelt. The two main protein fractions are gliadin (prolamin) and glutenin. These function in helping food maintain their shape by acting as a glue that is able to hold food together. It is not only contained in cereal proteins but also found in many other foods such as ice cream and sausages, it is also found in cosmetics (role as binder and filler). and in animal feed. It also gives foods their sticky-ness when it is added during processing. Without this protein rich mixture bread mixes would not hold together like they do or sausage would fall apart when made without gluten.

When cereals like wheat, rye, and barley are consumed, a hereditary autoimmune illness known as celiac disease can develop in the small intestine. Gluten ataxia is a neurological disorder that has symptoms including; behavioural change, weakness, tremor, ataxia and seizures. It is sometimes called gluten neuropathy or gluten encephalopathy. Non-celiac gluten sensitivity is a syndrome of people who experience an adverse reaction to the consumption of wheat-containing foods without evidence of celiac disease or wheat allergy.

There are no clear medical criteria for diagnosing this condition or determining its cause; however it may be genetic or related to the gut microbiome. Gluten sensitivity can cause the same clinical symptoms as celiac disease but without damage to the intestinal lining. In some cases, gluten sensitivity may also cause non-celiac gluten protein intolerance, a condition similar to celiac disease in which people experience symptoms following exposure to gluten but usually have no damage on their intestinal lining.

INTENDED USE

Gluten ELISA kit is a procedure intended for analysing quantitative concentrations of gliadin or gluten in foods (such as baby food, bakery products, meat, beer and chocolate). This assay has a minimum analytical sensitivity limit of 0.3 ppm.

CONTENT

All reagents supplied need to be stored at 2 °C – 8 °C, unopened reagents will retain reactivity until expiration date. Do not use reagents beyond this date.

Microtiter Plate: Coated with anti-gliadin.
Gliadin Standards 1-5: Concentration 0, 2, 6, 20, 60 ppm Gliadin.
Conjugate (Anti-Gliadin-Peroxidase).
Substrate Solution (TMB).
Stop Solution (0.5 M H2SO4).
Sample dilution buffer (Tris)(10x).
Washing Solution (PBS + Tween 20)(10x Concentrate).
Instruction Manual.

SENSITIVITY

The minimum detection sensitivity level of gluten or gliadin using current gluten ELISA kit was 0.3 ppm. The standard range for this assay is 2.0 – 60.0 ppm.

ASSAY CHARACTERISTICS

– Sensitivity: Limit of detection, LOD (0.03ppm), Limit of quantification, LOQ (0.2ppm)
– Cross Reactivity: Wheat (100%), Rye (100%), Triticale (40%), Barley (5%), Oats (< 0.003%). No cross reaction was detected for the following: Milk, Rice, Amaranth, Pork, Corn, Quinoa, Beef, Buckwheat, Teff, Egg, Millet, Cocoa, Soy.
– Intra-Assay Precision: 4 – 5%
– Inter-Assay Precision: 2 – 3%
– Linearity: 95 – 115%
– Recovery: Corn semolina (98%), Rice wafer (97%), Dark chocolate (97%), Beer (96%), Baby food (88%), Sausage (85%).

REFERENCES

Quantitative double antibody sandwich ELISA for the determination of gliadin. J Immunoassay Immunochem. (2012) 33 (4): 339-51. Gujral N., et al.
Application of Microwave Irradiation and Heat to Improve Gliadin-Detection and Ricin ELISA Throughput with Food Samples. Toxins (Basel). (2015) 7 (6): 2135-44. Garber E.A. and Thole J.
Monitoring of daily gliadin intake in patients on gluten-free diets. Prague Med Rep. (2011) 112 (1): 5-17. Gabrovská D., et al.
Recognition of gliadin and glutenin_fractions in four commercial glutenassays. J AOAC Int. (2010) 93 (1): 190-6. Allred L.K. and Ritter B.W.
Sensitive detection and quantification of gliadin contamination in gluten-free food with immunomagnetic beads based liposomal fluorescence immunoassay. Anal Chim Acta. (2013) 787: 246-53. Chu P.T. and Wen H.W.

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