Heterophile Antibody Test - Process

Process

The test relies on the agglutination of the horse RBCs by heterophile antibodies in patient's serum. Heterophile means it reacts with proteins across species lines. Heterophile also can mean that it is an antibody that reacts with antigens other than the antigen that stimulated it (an antibody that crossreacts). A 20% suspension of horse red cells is used in an isotonic 3-8% sodium citrate formulation. One drop of the patient’s serum to be tested is mixed on an opal glass slide with one drop of a particulate suspension of guinea-pig kidney stroma, and a suspension of beef red cell stroma; sera and suspensions are mixed with a wooden applicator 10 times. Ten micro liters of the horse red cell suspension are then added and mixed with each drop of adsorbed serum. The mixture is left undisturbed for one minute (not rocked or shaken). Examine for the presence or absence of red cell agglutination. If stronger with the sera adsorbed with guinea-pig kidney, the test is positive. If stronger with the sera adsorbed with beef red cell stroma, the test is negative. If agglutination is absent in both mixtures, the test is negative. A known 'positive' and 'negative' control serum is tested with each batch of test sera.

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