False negative and prozone reactions in tests for antibodies to Epstein‐Barr virus‐associated nuclear antigen

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Abstract
Titration of anti‐complementary human sera may yield false negative or prozone reactions in anti‐complement immunofluorescence (ACIF) tests for antibodies to Epstein‐Barr virus‐associated nuclear antigen (EBNA) when the human complement (C') is added to the serum dilutions prior to charging of the target cell smears (two‐stage ACIF test). This effect of anti‐complementary sera and with it a source of error is avoided by using a three‐stage ACIF technique; that is, when the target cell smears are successively overlayed with serum dilutions, C' and fluorescein isothiocyanate‐conjugated antibodies to human βC/βA globulins. The two‐ and three‐stage ACIF procedures yield comparable anti‐EBNA titers with non‐anti‐complementary sera.

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