What happens when the enzyme's action is not stopped within the specified time limits in an ELISA test?

In an ELISA (Enzyme-Linked Immunosorbent Assay) test, the enzyme used to detect the presence of an antigen or antibody in a sample produces a color change in the substrate when it catalyzes the reaction. This color change is directly proportional to the amount of enzyme present, which correlates with the target analyte in the sample.

When the action of the enzyme is not stopped within the specified time limits, it continues to react with the substrate, leading to an increased amount of the colored product. This excessive reaction can result in a falsely increased signal, meaning the final measurement will reflect a higher concentration of the analyte than is actually present in the sample. It is essential to adhere to the designated incubation times in an ELISA assay because properly controlling reaction conditions ensures accurate and reliable results.

In conclusion, not stopping the enzyme's action appropriately leads to a falsely increased result, which can compromise the test’s integrity and reliability.