title: Flow cytometry immunophenotyping in integrated diagnostics of patients with newly diagnosed cytopenia: one tube 10-color 14-antibody screening panel and 3-tube extensive panel for detection of MDS-related features.
abstract: Acute leukemia, myelodysplastic syndromes (MDS), myeloproliferative neoplasms and lymphomas are the most prevalent diagnoses in adults presenting with new onset cytopenia. Here, we describe two 10-color panels of surface markers (screening and comprehensive panel) applied at the Flow Cytometry Laboratory, University Health Network, Toronto, ON, Canada. A 10-color flow cytometry is applied using the stain-lyse-wash sample preparation method. In patients with <10% blasts and no clear involvement by hematological malignancy based on cytomorphological evaluation of bone marrow (BM) smear, the recently published one-tube 10-color 14-antibody screening panel is applied. This panel allows detection of major B- and T-cell abnormalities, enumeration of cells in blast region (CD45 dim), and gives insight into myeloid BM compartment, including calculation of four-parameter score for MDS-related abnormalities. In patients who present with ≥10 - <20% blasts in blood or BM smears, a comprehensive three-tube panel of surface markers is used up front. The analysis is focused on the detection of abnormal antigen expression patterns not seen in normal/reactive BM, according to the guidelines developed by International/European LeukemiaNet Working Group for Flow Cytometry in MDS. In patients with ≥20% blasts, an additional tube is added to allow the detection of cytoplasmic markers necessary to diagnose mixed phenotype acute leukemia.
authors.affiliation: Department of Pathobiology and Laboratory Medicine, University of Toronto, University Health Network, Toronto, ON, Canada.
authors.forename: A
authors.initials: A
authors.lastname: Porwit
authors.affiliation: Department of Pathobiology and Laboratory Medicine, University of Toronto, University Health Network, Toronto, ON, Canada.
authors.forename: A
authors.initials: A
authors.lastname: Rajab
mesh_terms: D000906:Antibodies / Q000276:immunology; D000954:Antigens, Surface / Q000276:immunology; D003116:Color; D005434:Flow Cytometry / Q000295:instrumentation / Q000379:methods*; D006801:Humans; D016130:Immunophenotyping / Q000379:methods*; D008214:Lymphocytes / Q000276:immunology / Q000378:metabolism / Q000473:pathology; D009190:Myelodysplastic Syndromes / Q000276:immunology / Q000378:metabolism* / Q000473:pathology; D010198:Pancytopenia / Q000276:immunology / Q000378:metabolism* / Q000473:pathology; D015203:Reproducibility of Results
chemical_list: D000906:Antibodies; D000954:Antigens, Surface
keywords: Flow cytometry; leukemia; lymphoma; myelodysplastic syndromes