Reducing ELISA detection errors In the case of coexistence of natural factors (reagent stability, accuracy, instrument precision) and human factors (operators), it is still a topic worth discussing to accurately report experimental results. Now discuss from the following aspects. (1) The inspection technician should have basic qualities and practical experience in laboratory operations. Ability to operate experimental instruments and instruments proficiently, with a certain ability to summarize and analyze problems, and timely and properly resolve unexpected situations in the experiment. (2) Use a calibrated micropipette to eliminate natural errors. The accuracy of the pipette is particularly important for quantitative detection. (3) Evaluate the practicality of reagents. The stability of the reagent is extremely important to the accuracy. Before the reagents are put into use, yin and yang controls and repeated comparison tests of samples should be conducted to determine whether the reagents meet the requirements before they can be used. (4) Read the manual carefully before operation. Perform standardized operations in strict accordance with the requirements of the manual. Reagents of different batches cannot be mixed. Where it is worthy of improvement, it can be improved only after it is established after repeated experiments, such as mastering the number of times of washing the board. (5) Add samples accurately and quickly. Chemical theory shows that the amount of product is directly related to the amount of reactant. The sample addition is inaccurate and the amount of enzyme product cannot be determined, which directly affects the color development results. In addition, the determination of the color depth and the value of A is related to the amount of color developer and stop solution added, so the sample should be added carefully. Experiments that require samples to be completed within a certain period of time. If the samples are added slowly, errors will occur, and the reagents will be exposed for a long time, especially when the room temperature is too high, the storage period will be shortened or even invalid. (6) Wash thoroughly. If the plate washing is not thorough, the background color of the enzyme conjugate will show a false positive. In addition, the washing liquid should be fresh, and it is used now. The old washing liquid will have a background increase, and false positives may also appear. (7) Strictly control the reaction time. If the reaction time is too long, the enzyme will be inactivated; if the reaction time is too short, the enzyme conjugate will not be able to fully bind with the microbial antigen and antibody in the serum. The product structure is loose and unsteady, and it is easy to wash off, which may cause false negatives. (8) Strictly master the color development time. The color development time is too short. After the termination of the addition of the termination solution, the amount of substrate conjugate is too small, and false negatives are likely to occur. Color development after the time required for color development should be judged as a false positive, which may be related to the reagent itself. Immediately after adding the coloring agent, the color can not be reported positive, which may be the result of background color development. (9) Pay attention to the storage period of reagents. Reagents beyond the storage period cannot be used.
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