A Fluorometric Assay for Red Blood Cell Antibodies

Schreiber, Alain A.B.; Lambermont, Micheline; Strosberg, A. D.; Wybran, Joseph

doi:10.1046/j.1537-2995.1981.21281178153.x

Cited by 7 publications

(1 citation statement)

References 8 publications

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“…It can be seen that there is still uncertainty about the sensitivity of the anti introduction The correlation of the strength of the anti globulin test with the number of IgG mole cules on the erythrocyte has previously been studied for a limited number of antibodies by radioimmunoassay [1][2][3][4], complement-fix ing antibody consumption assay [5,6], poly vinylpyrrolidone augmented automated as say [7], enzyme-linked assay [8], immunofluorimetric assay [9,10], and, more recent globulin test and the degree of correlation with the number of molecules present. Sim ilarly, there is uncertainty that the strength of the direct antiglobulin test (DAT) can always be taken as a reliable guide to the number of IgG molecules coating the cell, particularly in cases of auto-immune hae molytic anaemia (AIHA) where the DAT is negative [4][5][6]10].…”

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confidence: 99%

Quantitation of IgG on Erythrocytes: Correlation of Number of IgG Molecules per Cell with the Strength of the Direct and Indirect Antiglobulin Tests

Merry¹,

Thomson²,

Rawlinson³

et al. 1984

Vox Sanguinis

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The number of IgG molecules bound to the erythrocyte surface for a given agglutination score in the antiglobulin test was studied with several different examples of anti-D, anti-E, anti-c, anti-Kell, anti-Fy^a, anti-Jk^a and immune anti-A antisera. The serological scores show a significant correlation with the mean values for bound IgG molecules within a restricted range, although the number bound for a given score may vary by up to 20%. The limit of detection was 100-120 IgG molecules per cell and when over 1,000 were bound, the cells were completely agglutinated. Anti-Kell bound under low ionic strength saline conditions required a greater number of molecules for a given agglutination strength. The relatively low levels of bound IgG necessary to give strong agglutination make the direct antiglobulin test (DAT) less valuable for following the progress of auto-immune haemolytic anaemia (AIHA) than a quantitative test. The latter test does not, however, provide any additional information in AIHA cases with a negative DAT as in these the anaemia does not appear to be due simply to the number of bound IgG molecules. Detection of certain antibodies may not be achieved simply by increasing the sensitivity of the antiglobulin test when correctly performed.

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mentioning

confidence: 99%

Quantitation of IgG on Erythrocytes: Correlation of Number of IgG Molecules per Cell with the Strength of the Direct and Indirect Antiglobulin Tests

Merry¹,

Thomson²,

Rawlinson³

et al. 1984

Vox Sanguinis

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Detection of Red Blood Cell—Bound Immunoglobulin G by Flow Cytometry and its Application in the Diagnosis of Autoimmune Hemolytic Anemia

Wang¹,

Shi²,

Zhou³

et al. 2001

Int J Hematol

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Detection of autoantibodies to erythrocytes is of fundamental importance in the diagnosis of autoimmune hemolytic anemia (AIHA). The routinely used direct antiglobulin test (DAT) has the disadvantage of low sensitivity. In this study, we investigated the optimal test conditions of measurement of red blood cell (RBC)-bound immunoglobulin (Ig) G by flow cytometry (FCM). We studied 64 patients with AIHA, 30 anemic patients diagnosed with other diseases, and 36 healthy individuals. In 33 AIHA patients who were found to have RBC-bound IgG, both the mean fluorescence intensity (MFI) and percentage of fluorescence-activated RBCs were remarkably increased and results of both were considered positive. In the remaining 31 AIHA patients who had positive results for RBC-bound complement C3d, the MFI and the percentage of fluorescence-activated RBCs was also increased and 17 patients (54.8%) were considered to have a positive result by this method. In anemic patients with negative DATs the results of FCM were always negative. These results could be confirmed by enzyme-linked immunosorbent assay (ELISA), and the values obtained by FCM and ELISA corresponded to titration scores of the DAT. Additionally, in 3 of the other 8 patients who were suspected to have DAT-negative AIHA, RBC-bound IgG was detected by FCM and ELISA. Our investigation demonstrates that FCM is a precise, reliable, and sensitive method of detecting RBC-bound autoantibodies and could be used as a new routine diagnostic technique for AIHA and other immune hemolytic anemias.

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Radioimmunoassay for immunoglobulin G autoantibody on the surface of mouse erythrocytes

Jones

Lee

Skinner

et al. 1983

Clinical Immunology and Immunopathology

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A Fluorometric Assay for Red Blood Cell Antibodies

Cited by 7 publications

References 8 publications

Quantitation of IgG on Erythrocytes: Correlation of Number of IgG Molecules per Cell with the Strength of the Direct and Indirect Antiglobulin Tests

Quantitation of IgG on Erythrocytes: Correlation of Number of IgG Molecules per Cell with the Strength of the Direct and Indirect Antiglobulin Tests

Detection of Red Blood Cell—Bound Immunoglobulin G by Flow Cytometry and its Application in the Diagnosis of Autoimmune Hemolytic Anemia

Radioimmunoassay for immunoglobulin G autoantibody on the surface of mouse erythrocytes

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