Quantitation

The agglutination method has fallen somewhat into disuse for research and, to some extent, even for diagnosis because it is not possible to precisely quan-titate the reaction. Concentrations of agglutinating antibody are assayed by preparing a series of doubling dilutions of the test serum, adding cells to each dilution, incubating the cell suspension, and then observing the highest dilution of the serum which produces a visual agglutination pattern at the bottom of the tube. Thus, failure to observe the presence or absence of agglutination in one tube or one doubling dilution represents a 100% error in determining the quantity of antibody. Therefore, agglutination has been replaced by more quantifiable assays, including the ELISA, various radioimmunoassays and other types of immunoassays. However, agglutination can be quantitated. The actual amount of antibody nitrogen or protein bound by cells can be determined, but is too complicated and time-consuming to be a rou tine assay. Modern technology has provided a number of methods for the quantitation of agglutination through measurements of turbidity, light scattering or the mean diffusion constants of the particulate suspension.

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