GLIADORPHIN (GLIADOMORPHIN) ELISA KIT
INTENDED USE
This Gliadorphin (Gliadomorphin) ELISA Kit is intended for the determination of Gliadorphin in urine. It is for in vitro diagnostic use only.
INTRODUCTION
Gliadorphin is a 7 amino acids peptide which is formed during digestion of the gliadin component of the gluten protein. Gluten-derived peptides bind to opioid receptors in the brain and exhibit morphine-like effects, for example like heroin. These compounds have been shown to react with areas of the brain which are involved in speech and auditory integration.
Urine samples from people with autism, schizophrenia, and celiac disease contain high amounts of gliadorphin. It is suspected that this peptide may also be elevated in other disorders such as chronic fatigue, fibromyalgia, and depression. Symptom remission has been observed after exclusion of wheatand dairy products from the diet.
PRINCIPLE OF THE TEST
Gliadorphin (Gliadomorphin) ELISA Kit
This Gliadorphin (Gliadomorphin) ELISA Kit is intended for the determination of Gliadorphin in urine. It is for in vitro diagnostic use only. The Gliadorphin (Gliadomorphin) ELISA Kit is based on the method of competitive enzyme linked immunoassays.
Reaction buffer is used for sample preparation. Afterwards, the diluted samples and a Gliadorphin-derivative (tracer) are incubated in the microtiter plate wells coated with a Gliadorphin-antiserum. During the incubation, the target Gliadorphin in the sample competes with the tracer for the binding on the polyclonal antibodies immobilized on the wall of the microtiter wells. Gliadorphin in the sample displaces the tracer bound to the antibodies. Therefore, the concentration of the antibody-bound tracer is inverse proportional to the Gliadorphin concentration in the sample. During the second incubation step, a peroxidase-conjugated antibody, which binds to the tracer, is added to each microtiter well. After washing the unbound components, the peroxidase substrate tetramethylbenzidine (TMB) is added.
Finally, the enzymatic reaction is terminated by an acidic stop solution. The color changes from blue to yellow and the absorbance is measured in the photometer at 450 nm. The intensity of the yellow color is inverse proportional to the Gliadorphin concentration in the sample; this means high Gliadorphin concentration in the sample reduces the concentration of the antibody-bound tracer and lowers the photometric signal.
A dose response curve of absorbance unit (optical density, OD at 450 nm) vs. concentration is generated using the values obtained from the standard. Gliadorphin present in the patient samples is determined directly from this curve. The ELISA results are normalized to the creatinine concentration of the urine sample. For this reason, a parallel determination of the creatinine concentration is required.
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