Description

The Diagnostic Automation Inc. Gliadin IgG ELISA kit is designed to detect IgG class antibodies to Gliadin in human sera. Wells of plastic microwell strips are sensitized by passive absorption with Gliadin antigen. The test procedure involves three incubation steps: (1) Test sera (properly diluted) are incubated in antigen coated microwells. Any antigen specific antibody in the sample will bind to the immobilized antigen. The plate is washed to remove unbound antibody and other serum components. (2) Peroxidase Conjugated goat anti-human IgG (γ chain specific) is added to the wells and the plate is incubated. The Conjugate will react with Gliadin antibody immobilized on the solid phase in step 1. The wells are washed to remove unreacted Conjugate. (3) The microwells containing immobilized peroxidase Conjugate are incubated with peroxidase Substrate Solution. Hydrolysis of the Substrate by peroxidase produces a color change. After a period of time the reaction is stopped and the color intensity of the solution is measured photometrically. The color intensity of the solution depends upon the antibody concentration in the original test sample.

Quality control must be observed in using the MPO, Myeloperoxidase (p-ANCA) test. Each time the assay is run the Calibrator must be run in triplicate. A reagent blank, negative control and positive control must also be included in each assay. Calculate the mean of the three Calibrator wells. If any of the three values differ by more than 15% from the mean, discard that value and calculate the mean using the remaining two wells. The mean OD value for the Calibrator and the OD values for the Positive and Negative Controls should fall within the following ranges:



1. Negative Control < or = 0.250

2. Calibrator > or = 0.300

3. Positive Control > or = 0.500



Where:



a. The OD of the Negative Control divided by the mean OD of the Calibrator should be < or = 0.9.

b. The OD of the Positive Control divided by the mean OD of the Calibrator should be > or = 1.25.

c. If the above conditions are not met the test should be considered invalid and should be repeated.