Cytogenetics and flow cytometry in diagnosis of CLL – pro

Chronic lymphocytic leukemia displays a characteristic patterns of surface antigen expression (CD antigens), which facilitate their identification and proper classification and hence play an important role in instituting proper treatment plans. Multiparameter flow cytometric analysis has become commonplace in most laboratories for that purpose. Distinction between lymphoid and myeloid leukemias and of various subtypes of chronic lymphoproliferative disorders is crucially important. Several advances in flow cytometry, including availability of new monoclonal antibodies, improved gating strategies, and multiparameter analytic techniques, have all dramatically improved the utility of flow cytometry in the diagnosis and classification of leukemia.

Flow cytometery is very helpful in a diagnosis of any leukemic condition and can be diagnostic of CLL by showing CD5 positivity, especially when joined with CD19 and 20 in a multigated analysis. Flow cytometery represents one of several sources of information that go into making a secure diagnosis. It should not be relied on in isolation to make a diagnosis.

The role for cytogenetics once CLL is diagnosed in more complex. It can support the diagnosis by confirming abnormalities tyoical of CLL and also can provide prognostic information. A recent European guideline says: “Molecular cytogenetic analysis via fluorescent in situ hybridization (FISH) for the detection of unfavourable prognostic factors like deletions on chromosome 17p or 11q is recommended as a diagnostic procedure prior first-line treatment. The usage of unmutated VH status, VH 3.21 usage, ZAP70- and CD38 expression, serum markers like CD23, thymidine kinase and ß2-microglobuline for routine assessment is not advised. Further clinical trials are requested to standardize these prognostic markers and develop them to useful instruments in clinical practice.”

Salem DA, Stetler-Stevenson M. Clinical Flow-Cytometric Testing in Chronic Lymphocytic Leukemia. Methods Mol Biol. 2019;2032:311-321. doi: 10.1007/978-1-4939-9650-6_17. PMID: 31522426.

Rawstron AC, Kreuzer KA, Soosapilla A, et al. Reproducible diagnosis of chronic lymphocytic leukemia by flow cytometry: An European Research Initiative on CLL (ERIC) & European Society for Clinical Cell Analysis (ESCCA) Harmonisation project. Cytometry B Clin Cytom. 2018;94(1):121-128. doi:10.1002/cyto.b.21595

D’Archangelo M. Flow cytometry: new guidelines to support its clinical application.
Cytometry B Clin Cytom. 2007 May;72(3):209-10.

Eichhorst B, Dreyling M, Robak T, Montserrat E, Hallek M, ESMO Guidelines Working Group. Chronic lymphocytic leukemia: ESMO Clinical Practice Guidelines for diagnosis, treatment and follow-up. Ann Oncol. 2011 Sep. 22 Suppl 6:vi50-4

Hallek M, Cheson BD, Catovsky D, et al. Guidelines for the diagnosis and treatment of chronic lymphocytic leukemia: a report from the International Workshop on Chronic Lymphocytic Leukemia updating the National Cancer Institute-Working Group 1996 guidelines. Blood 2008; 111:5446.


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