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Appl Environ Microbiol. 1968 May; 16(5): 691-694
Copyright © 1968 American Society for Microbiology. All Rights Reserved.

An Improved Diluent for Rubella Hemagglutination and Hemagglutination-Inhibition Tests

Microbiological Associates, Inc., Bethesda, Maryland 20014
the Section on Infectious Diseases, National Institute of Neurological Diseases and Blindness, Bethesda, Maryland 20014

ABSTRACT

Rubella hemagglutinating (HA) antigen was prepared in BHK-21 tissue as 5% cell suspensions and from unconcentrated and 20x concentrated infected supernatant fluids. In some instances, unconcentrated fluids were treated with Tween 80 and ether; cell suspensions were treated with ether alone. Preparations were tested for HA activity in dextrose-gelatin-Veronal (DGV) buffer solutions; 0.85% NaCl; Sorenson's phosphate-buffered saline, pH 7.2; and a diluent of 0.9% NaCl, 0.1% CaCl₂ (anhydrous), and 0.1% MgSO₄·7H₂O. HA titers were consistently two- to fourfold higher in the saline with added Ca⁺⁺ and Mg⁺⁺ than in DGV. Hemagglutination-inhibition titers of paired human sera were the same in either diluent. It is suggested that the interaction between rubella HA antigen and the red cells of young (less than 1-day-old) chicks may be at least partially ion dependent and that titers are enhanced by increased quantities of divalent cations.