Hemagglutination techniques

Dewdney J (1980) Pseudo-allergic reactions to antibiotics. In Dukor P et al. (eds) PAR Pseudo-allergic reactions, vol 1. Karger, Basel, p 273 De Week AL (1964) Penicillin allergy, its detection by an improved hemagglutination technique. Nature 202 975... 

Additional serological techniques, such as complement consumption tests, radioimmunoprecipitation (Gleich and Stankievic 1969), fluorescence polarization (Dandliker et al. 1965), and variations of sandwich hemagglutination techniques (e.g., red-cell-linked antigen-antiglobulin reaction Kraft et al. 1976), have been used in the detection of antipenicillin antibodies, but all appear to have been supplanted by modern RIA and ELISA techniques. 

The first description of antibodies against polynucleotides was that of the anti-DNA antibody. Such antibodies, in sera of patients, were demonstrated by precipitation, by complement fixation, and by hemagglutination techniques (Seligmann, 1957 Ceppelini et al., 1957 Seligmann and Milgrom, 1957 Robbins et al., 1957). 

Forerunners of nonisotopic immunoassay had already appeared before radioimmunoassay was developed. For example, nephelometry is based on precipitation, which is known as the classical immune reaction, and the ideas of particle immunoassay and viroimmunoassay seem to have developed from the hemagglutination test. The principles of enzyme and fluorescence immunoassay had already been used as enzyme and fluorescence antibody techniques in histochemical analysis. In 1971, two groups reported use of an enzyme immunoassay (E5, V2). Leute et al. reported spin immunoassay, which has spurred recent development of nonisotopic immunoassays (L5). 

A sample of virus should be retained and tested by hemagglutination on chicken or duck red blood cells. An excellent description of this technique has been published (7). 

In addition to the direct reaction of the antigen with antibody, techniques are available for performing indirect reactions. Antigens can be coupled chemically or via a tannic acid procedure to red blood cells. The antigen-coated erythrocytes in the presence of specific antibody then agglutinate (clump) as do red blood cells in the presence of antibody to the red blood cells (hemagglutination). 

This procedure is of great potential utility for the identification and enumeration of cells that secrete an antigenic product. Patterned after the assay just described, it employs similar techniques with two major differences The coating for the indicator red cells consists of purified antibody, as in the reverse passive hemagglutination previously described, and an antiserum against the secretion product is used as the developing agent. 

In indirect or passive hemagglutination, the erythrocytes are used as a particulate carrier of foreign antigen (and in some tests of antibodies) this technique has wide applications. Other materials available in the form of fine particles, such as bentonite and latex, also have been used as antigen carriers, but they are more difficult to coat, standardize, and store. In a related variation of tliis technique, known as hemagglutination inhibition, the ability of antigens, haptens, or other substances to specifically inhibit hemagglutination of sensitized (coated) cells by antibody is determined. 

Tg antibodies are directed against the Tg protein, a major constituent of thyroid colloid. Several different techniques have been used to detect and quantify TgAb in peripheral blood. These include passive hemagglutination, the agar gel diffusion precipitin technique, immunofluorescence of tissue sections, enzyme-linked immunosorbent assay (ELISA), radioassay techniques, and chemiluminescence-based immunometric assays. 

Wide and his colleagues (W5, W7) were the first to apply the technique of hemagglutination-inhibition to the estimation of urinary LH. They found that some antisera raised against HCG were incapable of distinguishing between HCG and LH, and, accordingly, they were able to establish an assay system for LH using an antiserum raised to HCG and HCG-coated red blood cells. Taymor (T2) used a similar system to assay human urinary LH. He also employed an HCG antiserum and latex particles coated with this hormone the system was specific in that a cross reaction with ovine LH was not observed however, its specificity with respect to FSH was not reported. Taymor (T2) found it necessary to extract LH from urine prior to immunoassay. For this purpose he preferred precipitation with acetone to treatment with alcohol, since trace amounts of the latter interfered with the antigen-antibody reaction. 

Precision. Figures for the index of precision (A) using such techniques do not yet appear to have been calculated. According to Stockell and Hartree (S25), the precision of hemagglutination-inhibition tests is reasonably satisfactory in skilled hands, the fiducial limits of... 

Practicability. The radioimmunoassay is easy to perform, and unlike the hemagglutination-inhibition technique, requires only small amounts of purified gonadotropic hormones. The main disadvantage of radioimmunoassays is that they require costly equipment and laboratories which are specially constructed for the handling of relatively large quantities of isotopic iodine. 

Passive hemagglutination has a lower detection limit of 3-6 ng antibodies per ml serum. In this technique, soluble antigens are bound to the surface of erythrocytes, which are agglutinated when the antigen-antibody reaction occurs. 

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