Question

A person infected with HIV was tested with the HIV ELISA as described in lecture. The technician who performed the ELISA made a mistake and forgot to wash the wells after addition of the primary antibodies. Note: all other steps were performed correctly. Describe what would happen in the test wells during this faulty ELISA procedure and include the final test results produced.

Answer 1

In ELISA, the washing step is important to wash the unbound antibodies and washing will reduce any non-specific binding that may occur.

If the technician forgot to wash the test well after the addition of primary antibodies then all the unbounded antibodies will not be washed out and which will result in high background signal. There will be poor replicate data in ELISA, the results of ELISA will be inconsistent from assay to assay and there will be the large coefficient of variation.

Eventually, the test results will be possibly indeterminate.