You are provided with the following:

  • Two serum samples, one from Mr Campbell, and one from the kidney donor
  • Alkaline phosphatase-conjugated rabbit anti-human IgG and IgM antibodies
  • 96 well ELISA plate pre-coated with CMV antigen
  • CMV-positive control serum
  • CMV-negative control serum
  • Enzyme substrate (p-nitrophenyl phosphate)
  • Phosphate buffered saline (PBS)
  • 3M NaOH

The ELISA technique is a solid-phase serological assay which can be used to detect, and if appropriate standards are run, to quantify any known antigen or antibody. The virtues of this assay are that:

  1. It is very sensitive: the enzyme molecule linked to the detection antibody provides an amplification of the signal
  2. Large numbers of samples can be examined (with or without automation)
  3. Results can be obtained within hours

The starting point is to coat a solid phase support, e.g. a plastic plate, with either a specific antigen (if antibody is to be measured - as in this case) or antibody (to measure an antigen). Subsequently, a binding step with the target molecule is performed, followed by a detection step with a derivatized antibody linked to an enzyme molecule such as alkaline phosphatase. The enzyme converts a colourless substrate to a coloured product.

In this assay, you will measure the amount of CMV-specific IgG and IgM in the sera.

Start the assay

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