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Measurement of Rubella Antibody by Hemagglutination Inhibition

II. Characteristics of an Improved HAI Test Employing a New Method for the Removal of Non-Immunoglobulin HA Inhibitors from Serum¹

From the Department of Epidemiology and Public Health and the Department of Microbiology, Yale University School of Medicine, New Haven, Connecticut

Abstract

A method employing dextran sulfate and CaCl₂ (DS-C) for removing nonspecific inhibitors of rubella virus hemagglutinin from test sera has been described. Sera treated by this method were shown to be free of detectable inhibitor. No quantitative changes in IgG, IgA and IgM could be detected, nor was any qualitative change in IgG observed as a result of DS-C treatment. The hemagglutination-inhibition (HAI) test employing DS-C treated sera was shown to be a true measure of actual rubella antibody content, and 100% of the HAI titers determined on 47 paired sera were identical or equivalent (±1 dilution).

Kaolin by comparison was shown to be an unreliable means of specifically removing non-immunoglobulin inhibitors. Kaolin failed to remove inhibitors completely from many sera, and it sometimes removed specific antibody from others.

A modified heparin-MnCl₂ procedure was found to give erratic and unreliable results. The basis for these undesirable characteristics of the heparin-MnCl₂ method was explored and an improved heparin-MnCl₂ procedure was tried. This procedure gives reproducible and reliable results, but has the disadvantage that it fails to remove all traces of inhibitor from some sera, thus giving rise to false positive tests.

Footnotes

This work was supported by Contract PH-43-68-1037 from the National Institute of Allergy and Infectious Diseases, United States Public Health Service.