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A systematic study was made of certain variables of the rubella hemagglutination-inhibition (HI) test system and their effect on antigen and antibody titers. Erythrocytes from pigeons and 1-day-old chicks gave similar antigen and antibody titers, but goose erythrocytes gave lower titers. Indicator erythrocytes could be stored in Alsever's solution at 4 C for as long as 2 weeks without losing sensitivity in hemagglutination (HA) and HI tests. Antigen titers varied by eightfold or more in different diluent systems; titers were generally higher atp H 6.2 than atp H 7.2. A diluent without Ca2+ gave antigen titers as high as those obtained in diluents with added Ca2+ ions. Antibody titers also varied in different diluent systems. HEPES diluents atp H 6.2 gave higher antibody titers than those obtained in other diluents, but occasional “false-positive” inhibition reactions were seen. Kaolin suspended in borate saline atp H 9.0 effectively removed inhibitor from sera without absorbing specific antibody, but atp H 7.3 it removed various amounts of specific antibody. Antibody titers of sera treated with kaolin atp H 9.0 were similar to those of sera treated with heparin-MnCl2 ; treatment with dextran sulfate-CaCl2 gave lower antibody titers. Antigens varied widely in sensitivity for detecting HI antibody and in the ability to detect diagnostically significant increases in antibody. Sensitivity in detecting antibody was not related to the HA titer of the antigens. Tween-ether-treated antigens gave lower antibody titers but were more reliable than corresponding untreated antigens for serological diagnosis of infection.

Variables of the Rubella Hemagglutination-Inhibition Test System and Their Effect on Antigen and Antibody Titers