An automatable format for accurate immunohematology testing by flow cytometry.
BACKGROUND
Current immunohematology testing methods have limitations including cost, throughput, and adaptability to automation. Furthermore, current automated and semiautomated workstations cannot accommodate many other tests relevant to blood transfusion.


METHODS
Authentic clinical samples from hospitalized patients were tested for ABO group, D type, and presence of RBC alloantibodies by column agglutination technology (CAT), standard tube methods, and a recently developed flow cytometry (FC) technique. Included were challenging samples with rouleaux, autoantibodies, mixed-field reactions, and weak antibodies. Antibody staining of RBCs for FC was initially performed in test tubes and subsequently in microtiter filter plates interfaced with a vacuum manifold.


RESULTS
When antibody staining was performed in tubes, FC testing determined the correct ABO group and D type for 99.1 percent of 222 clinical samples, as compared to accuracies of 91.9 percent for CAT and 95.0 percent for standard tube testing. FC testing also detected 99.5 percent of clinically relevant RBC alloantibodies in 239 patient samples, as compared to 98.9 percent for CAT and 94.7 percent for LISS-IAT. Using the FC filter plate technique, 104 of 109 samples (95.4%) were correctly typed for ABO and D (the remaining five samples were read as "no type determined" due to RBC and serum testing discrepancies), and RBC alloantibodies of the IgG and IgM classes were correctly identified in 98.3 percent of samples.


CONCLUSIONS
Optimized FC testing methods that are comparable in accuracy to standard CAT and tube methods are described. When used with filter plates, this methodology should allow rapid and cost-effective immunohematology testing of both patient and donor samples in an automated workstation format. The same workstation should support automation of other pretransfusion assays that can be analyzed by FC.
Roback
John D
JD
Transfusion Medicine Program, Department of Pathology and Laboratory Medicine, Emory University School of Medicine, Atlanta, Georgia, USA. jroback@emory.edu
Barclay
Sheilagh
S
Hillyer
Christopher D
CD
D000017:ABO Blood-Group System; D001322:Autoanalysis; D001323:Autoantibodies / Q000097:blood; D001788:Blood Grouping and Crossmatching / Q000379:methods*; D004912:Erythrocytes / Q000276:immunology; D005434:Flow Cytometry / Q000379:methods; D006801:Humans; D007074:Immunoglobulin G / Q000097:blood; D007075:Immunoglobulin M / Q000097:blood; D007518:Isoantibodies / Q000097:blood; D012204:Rh-Hr Blood-Group System
D000017:ABO Blood-Group System; D001323:Autoantibodies; D007074:Immunoglobulin G; D007075:Immunoglobulin M; D007518:Isoantibodies; D012204:Rh-Hr Blood-Group System