Leukemia/Lymphoma Phenotyping Evaluation by Flow Cytometry

Last Literature Review: June 2019 Last Update:

Leukemia and lymphoma analysis by flow cytometry aids in identifying the tumor lineage, which in most cases is identified as T cell, B cell, or myeloid. Lineage identification can provide a confirmatory diagnosis or differential diagnosis, prognosis, and treatment options.

Disease Overview

Diagnosis/Treatment Issues

  • Phenotyping by flow cytometry can aid in the evaluation of hematopoietic neoplasms.
    • Specimens include bone marrow, whole blood, tissue, or fluid.
  • Phenotyping may aid in monitoring response to therapy in individuals with an established diagnosis of hematopoietic neoplasms.

Test Interpretation

  • Markers are analyzed as needed, based on clinical evidence, to fully characterize any abnormalities identified by the screening panel.
    • Additional markers are selected based on pathologist interpretation of the screening panel results.
  • Antigens included:
    • T cell: CD1a, CD2, CD3, CD4, CD5, CD7, CD8, TCR γ-δ, cytoplasmic CD3
    • B cell: CD10, CD19, CD20, CD22, CD23, CD103, CD200, kappa, lambda, cytoplasmic kappa, cytoplasmic lambda
    • Myeloid/monocyte: CD11b, CD13, CD14 (Mo2), CD14 (MY4), CD15, CD33, CD64, CD117, myeloperoxidase
  • Miscellaneous: CD11c, CD16, CD25, CD30, CD34, CD38, CD41, CD42b, CD45, CD56, CD57, CD61, HLA-DR, glycophorin, TdT, bcl-2, ALK-1, CD123, CD138, CD200, CD26, CD45, CRLF-2

Clinical Sensitivity

Limit of detection is 0.011.0% depending on phenotype and disease.


  • Antigens will be reported as positive or negative.
  • Interpretive comments that further characterize intensity patterns are included.
    • Dim, bright, variable, or partial may be reported.
  • Light-chain expression may be reported as polytypic/polyclonal or restricted/monotypic/monoclonal.
    • May include kappa/lambda ratio.
  • Pattern of CD antigen testing will be interpreted with recommendations for further testing, if indicated.


  • Some hematopoietic neoplasms do not show phenotypic abnormalities and may not be detected by flow cytometry.
  • Poor cell viability may adversely affect antigens and impede the ability to properly identify neoplastic cells.
  • Flow results cannot be used alone to diagnose malignancy.
    • Results should be interpreted in conjunction with morphology, clinical information, and other necessary ancillary tests for a definitive diagnosis.

Related Testing Strategy Information

Leukemia/Lymphoma ARUP Consult Resource

Acute leukemia

Follicular, Burkitt, or diffuse large B-cell lymphoma

Chronic lymphocytic leukemia, small lymphocytic lymphoma, hairy cell leukemia, and mantle cell lymphoma