A Flow Cytometric Study of Reagent Cells to Resolve ABO Typing Discrepancy

Abstract

Objectives RBC alloantibodies can lead to ABO grouping discrepancies unrelated to A or B antigens or antibodies posing challenges in the blood bank testing. Routine blood bank testing and flow cytometry were used to immunophenotype reagent cells and elucidate the cause of ABO discrepancies in two patients. Methods ABO discrepancy was identified in two patients after transfusion with several units of RBCs. For both patients, the pretransfusion type and screen demonstrated blood group A. Eight and 16 days later, both patients showed an apparent antibody to reagent group A cells, which prompted additional study with patients’ samples and flow cytometric testing of commercial reagent cells. Results In both patients’ specimens, posttransfusion evaluation demonstrated an emerging antibody to the Kell antigen (K). The RBCs of both patients typed negative for K, and both were transfused with K-positive RBCs. Flow cytometric analysis of reagent RBCs demonstrated that five of seven lot numbers were positive for K. Conclusions Emerging anti-K antibody led to agglutination of the K-positive reagent A1 cells, highlighting the importance of considering RBC alloantibodies and the composition of reagent cells when interpreting cases with an apparent ABO grouping discrepancy.

Blood Group Testing

“…A flow cytometry-based method has been established to detect the human ABO group and D type. It has higher specificity, sensitivity, and reproducibility compared to traditional hemagglutination methods and can easily detect immunoglobulin subtypes in addition to quantifying the antibodies (80)(81)(82)(83)(84). Flow cytometry is capable of detecting tiny subpopulations as an alternative and supplemental approach when blood groups cannot be detected by serology and results can be delivered in less than an hour (85).…”