title: Novel lymphocyte screening tube using dried monoclonal antibody reagents.
abstract: We previously developed a 10-color 11-antibody combination including a viability dye, to screen T-, B-, and natural killer (NK)-cell populations in blood, bone marrow, tissue, and body fluids. Recently, Beckman Coulter has introduced a line of dried reagents that, unlike liquid reagents and cocktails, require no refrigeration, titration, or manipulation before using. We evaluated custom tubes based on our standard lymphocyte screening panel, focusing on comparative analysis, ease of use, and advantages compared with our liquid reagent set. We tested 42 samples from blood (n = 15), bone marrow (n = 17), and tissue (n = 10) with the combination CD4/CD8/KAPPA/LAMBDA/CD19/CD56/CD5/CD20/CD10/CD3/CD45 and a vital dye by both methods and compared positivity and staining intensity for each antigen. Of the 42 samples, 5 were normal samples, 3 were red cell disorders, 20 were B-cell malignancies, 5 T-cell malignancies, 4 myeloid malignancies, and the remaining 5 were other diagnoses. Dried reagents gave equivalent staining intensity results to our standard panel in a variety of sample types, with diagnoses including reactive lymphocytosis, chronic lymphocytic leukemia, and various lymphomas. Our standard panel for evaluation of mature lymphoid malignancies allows rapid assessment of any sample type while providing direct assessment of viability. The dried reagent tube reduces preanalytical work, with simple addition of sample and the viability dye to the tube, saving time, reducing potential errors, and obviating need to titrate and monitor individual antibodies. With a shelf life of at least 12 months, the reagents also offer potential savings in reagent costs by reducing wastage due to expiration or tandem breakdown in standard liquid formulation.
authors.affiliation: Pathology and Laboratory Medicine, Division of Hematology, London Health Sciences Centre, London, Ontario, N6K 5W9, Canada.
authors.forename: Benjamin D
authors.initials: BD
authors.lastname: Hedley
authors.affiliation: Pathology and Laboratory Medicine, Division of Hematology, London Health Sciences Centre, London, Ontario, N6K 5W9, Canada.
authors.forename: Michael
authors.initials: M
authors.lastname: Keeney
authors.affiliation: Pathology and Laboratory Medicine, Division of Hematology, London Health Sciences Centre, London, Ontario, N6K 5W9, Canada.
authors.forename: Janice
authors.initials: J
authors.lastname: Popma
authors.affiliation: Department of Medicine, Division of Hematology, London Health Sciences Centre, Schulich School of Medicine Western University of Ontario, London, Ontario, N6K 5W9, Canada.
authors.forename: Ian
authors.initials: I
authors.lastname: Chin-Yee
mesh_terms: D000911:Antibodies, Monoclonal / Q000276:immunology*; D015703:Antigens, CD / Q000276:immunology; D005434:Flow Cytometry / Q000379:methods*; D006801:Humans; D016130:Immunophenotyping / Q000379:methods*; D007938:Leukemia / Q000276:immunology / Q000473:pathology; D008214:Lymphocytes / Q000276:immunology / Q000473:pathology*; D008218:Lymphocytosis / Q000175:diagnosis / Q000276:immunology / Q000473:pathology; D008223:Lymphoma / Q000175:diagnosis / Q000276:immunology / Q000473:pathology
chemical_list: D000911:Antibodies, Monoclonal; D015703:Antigens, CD
keywords: acute leukemia; duraclone; flow cytometry; immunophenotyping; screening tube