The Elisa test is an experimental diagnostic method with high sensitivity, strong specificity and good repeatability. Due to its stability, easy storage, easy operation, and objective judgment of results, it has been widely used in various fields of immunological testing. The ELISA detection kit is used for qualitative detection of anti-human hepatitis E (HEV) virus IgM antibody in human serum or plasma in vitro by ELISA. Method 1: Double antibody sandwich method for detection of unknown antigens: 1. Coating: Dilute the antibody to a protein content of 1-10 μg / ml with 0.05MPH9. Carbonate coating buffer. Add 0.1ml to the reaction well of each polystyrene plate, overnight at 4 ℃. The next day, the solution in the well was discarded and washed 3 times with washing buffer for 3 minutes each time. (Referred to as washing, the same below). 2. Add sample: add 0.1ml of the diluted test sample to the above-mentioned coated wells and incubate at 37 ℃ for 1 hour. Then wash. (Make blank wells, negative control wells and positive control wells at the same time). 3. Add enzyme-labeled antibody: add 0.1ml of freshly diluted enzyme-labeled antibody (dilution after titration) to each reaction well. Incubate at 37 ° C for 0.5 to 1 hour and wash. 4. Add substrate to develop color: add 0.1ml of temporarily prepared TMB substrate solution to each reaction well at 37 ℃ for 10-30 minutes. 5. Terminate the reaction: Add 0.05ml of 2M sulfuric acid to each reaction well. 6. Judgment of results: The results can be directly observed with the naked eye on a white background: the darker the color in the reaction well, the stronger the positive degree, and the negative reaction is colorless or extremely light. "-" Indicates. OD value can also be measured: on ELISA detector, at 450nm (410nm if ABTS color is developed), the OD value of each well is measured after zero adjustment with a blank control well, if it is greater than 2.1 times the OD value of the specified negative control , That is positive. Method 2: Indirect method for detecting unknown antibodies: Dilute the known antigen to 1 ～ 10μg / ml in coating buffer, add 0.1ml per well, and overnight at 4 ℃. Wash 3 times the next day. Add 0.1ml of the diluted test sample (unknown antibody) to the coated wells, incubate at 37 ℃ for 1 hour, and wash. (Make blank, negative and positive well control at the same time) Add 0.1ml of fresh diluted enzyme-labeled secondary antibody (anti-antibody) to the reaction well, incubate at 37 ℃ for 30-60 minutes, wash, and wash with DDW one last time. The remaining steps are the same as 4, 5 and 6 of the "double antibody sandwich method".
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