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 ofMDS‐related features

Abstract

Summary 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.

Blastic plasmacytoid dendritic cell neoplasm with leukemic presentation: 10‐Color flow cytometry diagnosis and HyperCVAD therapy

“…Multicolor 10-color FCM immunophenotypic analysis with University Health Network AML panel [13] was performed on bone marrow aspirate samples collected in EDTA from six patients. The neoplastic cells were gated using the right angle light scatter and CD45 expression.…”

Initial flow cytometric evaluation of the Clearllab lymphoid screen

“…As the incidence of hematological malignancies, including leukemia and lymphoma (L&L) continues to increase (2,3) FCI remains an indispensable tool for diagnosing and monitoring these patients (4). It now requires fewer tubes to perform complex testing on small samples, and in some cases more rapid and effective immunophenotyping of samples in high volume laboratories (5)(6)(7)(8). It now requires fewer tubes to perform complex testing on small samples, and in some cases more rapid and effective immunophenotyping of samples in high volume laboratories (5)(6)(7)(8).…”

Aligning the flow cytometric evaluation with the diagnostic need: an evidence-based approach

“…The 10-colour, 14-antibody screen used by Toronto General Hospital focuses on the detection of abnormal antigen patterns not seen in normal/reactive bone marrows, and has been validated for screening for acute leukaemia, lymphoma and myelodysplastic syndrome10 11 We agree that a thorough validation process which includes the development and training of staff to use standardised analysis templates designed to clearly separate normal from abnormal populations is key to a successful implementation. Our laboratory also plans to incorporate key features of the Toyota Production System in order to facilitate a lean workflow.…”