An evaluation of a volumetric method for the flow cytometric determination of residual leukocytes in blood transfusion units

Abstract

The removal of leukocytes from blood components helps to prevent or reduce some adverse reactions that occur after blood transfusions. The implementation of the leukodepletion process in the preparation of blood units requires quality control, consisting of a reliable cell counting method to determine residual leukocytes in blood components. The most widely used methodology is a flow cytometric bead-based counting method. To avoid the need for commercial counting beads, we evaluated a volumetric counting method of leukocyte enumeration. A total of 160 specimens of leukodepleted plasma, red cell and platelet units, as well as 58 samples of commercially available controls containing different concentration levels of leukocytes, were included in the study. The conventional quality control method using the bead-based counting method performed with the FACSCalibur flow cytometer was compared to the bead-based counting method and the volumetric counting method performed with the MACSQuant 10 flow cytometer. Our results show that the MACSQuant bead-based method, as well as the volumetric MACSQuant method, meet the sensitivity requirements of residual leukocyte enumeration when compared to the gold standard, bead-based FACSCalibur method. We conclude that the volumetric method can be a substitute for the bead-based counting of residual leukocytes in a variety of blood components.