Flow-Cytometric Quantitation of Anti-D Antibodies
Article first published online: 1 MAY 2003
Volume 72, Issue 3, pages 172–176, April 1997
How to Cite
Hildén, J.-O., Backteman, K., Nilsson, J. and Ernerudh, J. (1997), Flow-Cytometric Quantitation of Anti-D Antibodies. Vox Sanguinis, 72: 172–176. doi: 10.1046/j.1423-0410.1997.7230172.x
- Issue published online: 1 MAY 2003
- Article first published online: 1 MAY 2003
Objectives: Quantitation of Rh antibodies is important clinically in predicting the risk of hemolytic disease of the newborn. We describe a flow cytometry method for the quantitation of anti-D antibodies that we developed in parallel to a recently described method. Methods: As a secondary antibody we used whole IgG instead of Fab molecules. The advantages, besides lower cost, include a stronge fluorescence signal with no need for amplification, and the possibility of diluting samples to minimize the risk of agglutination by IgM antibodies. We did extensive studies on reproducibility. Results: Reproducibility was superior to the autoanalyzer method. The two methods were roughly in agreement in estimating low, medium, or high levels of anti-D with a correlation coefficient of 0.89. The autoanalyzer measures the in vitro agglutination of all anti-D antibodies whereas flow cytometry measures the amount of IgG anti-D bound to red cells, which is more like the in vivo situation. Conclusion: Further studies in a clinical setting will show whether flow-cytometric quantitation may improve the diagnostic value of anti-D concentration measurement.