RESUMO
This paper describes an enzyme immunoassay for the quantitative determination of IgG, IgA, and IgM immunoglobulins on RBCs. Ether eluates made from RBCs were followed by an enzyme-linked immunosorbent assay of immunoglobulin concentration. Calibration curves were derived from immunoglobulin standards and the number of molecules of each isotype per RBC was calculated. The assay was carried out in 200 healthy blood donors and 62 patients with warm autoimmune hemolytic anemia (AIHA), two of them with a negative DAT. For healthy blood donors, mean values were 58 IgG, 16 IgA, and 3 IgM molecules per RBC. For patients with a positive DAT, the mean values were 3435 IgG, 157 IgA, and 69 IgM molecules per RBC. An increased level of IgA was found in 12 patients without IgA autoantibodies demonstrable in RBC eluates. Increased IgG levels were also observed in patients with a negative DAT and, in one case, an increased level of IgA was also found. The enzyme-linked immunosorbent assay using ether eluates is a sensitive method for quantitating RBC autoantibodies in patients with AIHA as well as immunoglobulins bound to RBCs in healthy individuals.
RESUMO
The efficacy of different methods for anti-red cell antibodies detection was assessed, variations being found in accordance with the specificity of the alloantibodies. The usefulness of enzyme tests in anti-Rh antibody detection was demonstrated, as well as that of low ionic strength saline solutions in detecting anti-Kell, anti-Duffy and anti-Kidd antibodies. Serum precipitation with 15% polyethyleneglycol 8000 previously to indirect antiglobulin test was found the most sensitive method, providing the best results in all the antibodies studied.