What are the steps of the complement fixation test?

The steps of the complement fixation test are described below.

1. The serum sample is first heated to about 56°C to eliminate existing complement proteins. 
2. Treat the serum sample with washed sheep red blood cells (RBCs) to prevent interference from cross-reactive anti-RBC antibodies. 
3. Introduce the antigen and complement into the sample.
4. Incubate the mixture at 37°C for 30 minutes, providing an optimal environment for the formation of antigen-antibody complexes
5. Add an indicator and observe the sample for any alterations, indicating either the occurrence or absence of hemolysis.