Serum level of total IgE:
Quantitative measurement of serum level of total IgE was done using a commercially available sandwich enzyme-linked immunosorbent assay (ELISA) Kit supplied by Chemux Bioscience, Inc. (CA, USA, Cat No.10602) according to the manufacturer’s instructions. The results were expressed in IU/mL. The total IgE level in a normal, allergy-free adult is less than 150 IU/ml of serum. The minimum detectable concentration of IgE by this assay is estimated to be 5.0 IU / ml. The Absorbance of standards and samples were measured at 450 nm using a microtiter plate ELISA reader (Biotek, USA).
Measurement of serumzonulin level:
Quantitative measurement of serum level of zonulin was done using a commercially available sandwich enzyme-linked immunosorbent assay (ELISA) Kit supplied by MyBiosource, Inc. (USA, Cat No: MBS774026) according to the manufacturer’s instructions. The results were expressed in ng/ml. Absorbance of standards and samples were measured at 450 nm using a microtiter plate ELISA reader(Biotek, USA). The kit detection range was 20ng/ml - 800ng/ml.