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HelpMayo Test ID SZDIA Sezary Diagnostic Flow Cytometry, Blood
Ordering Guidance
This test is not indicated for monitoring patients with a diagnosis of Sezary syndrome. For monitoring purposes, order SZMON / Sezary Monitoring Flow Cytometry, Blood.
Specimen Required
Container/Tube:
Preferred: Yellow top (ACD solution A or B)
Acceptable: Lavender top (EDTA), green top (sodium heparin)
Specimen Volume: 6 mL
Collection Instructions:
1. Send whole blood specimen in original tube. Do not aliquot.
2. Label specimen as blood.
Useful For
Identifying immunophenotypically aberrant T-cell populations with restricted expression of T-cell receptor beta-chain constant in peripheral blood, to roughly assess the circulating tumor burden in cutaneous T-cell lymphomas
Additional Tests
| Test ID | Reporting Name | Available Separately | Always Performed |
|---|---|---|---|
| FIRST | Flow Cytometry, Cell Surface, First | No | Yes |
| ADD1 | Flow Cytometry, Cell Surface, Addl | No | Yes |
Reflex Tests
| Test ID | Reporting Name | Available Separately | Always Performed |
|---|---|---|---|
| FCIMS | Flow Cytometry Interp, 9-15 Markers | No | No |
| FCINS | Flow Cytometry Interp,16 or greater | No | No |
Testing Algorithm
This Sezary panel is ordered for patients with a clinical suspicion of Sezary syndrome or cutaneous T-cell lymphoma with peripheral blood involvement without a previously confirmed diagnosis. A Triage panel and Sezary panel will always be performed. This test is not indicated for monitoring peripheral blood involvement in patients with a diagnosis of Sezary syndrome or mycosis fungoides. For monitoring purposes, order SZMON / Sezary Monitoring Flow Cytometry, Blood.
The panel is charged based on number of markers tested (FIRST for first marker, ADD1 for each additional marker). In addition, reflex testing may occur to fully characterize a disease state or clarify any abnormalities from the screening test. Reflex tests will be performed at an additional charge for each marker tested (ADD1 if applicable).
The testing process begins with a triage panel to evaluate for and exclude monotypic B cells or increased numbers of blasts. The triage panel also includes antibodies to assess the number of CD3-positive T cells and CD16-positive/CD3-negative natural killer cells present. Additional reflex testing may occur to fully characterize a disease state per algorithmic approach or clarify any abnormalities from the screening test at an additional charge for each marker tested (ADD1 if applicable). For a comprehensive list of potential additional panels, refer to LCMS / Leukemia/Lymphoma Immunophenotyping, Flow Cytometry, Varies.
The Sezary panel will further evaluate the T cells for expression of CD2, CD4, CD5, CD7, CD8, CD26, and TRBC1.
Method Name
Immunophenotyping
Reporting Name
Sezary Diagnostic Flow Cytometry, BSpecimen Type
Whole bloodSpecimen Minimum Volume
1 mL
Specimen Stability Information
| Specimen Type | Temperature | Time | Special Container |
|---|---|---|---|
| Whole blood | Ambient (preferred) | 4 days | |
| Refrigerated | 4 days |
Reject Due To
| Gross hemolysis | Reject |
| Gross lipemia | OK |
Clinical Information
Sezary syndrome (SS) and mycosis fungoides (MF) are two distinct but intimately related T-cell lymphoproliferative disorders involving the skin and are commonly referred to as cutaneous T-cell lymphomas (CTCLs). SS is defined by the triad of erythroderma, generalized lymphadenopathy, and the presence of circulating cells with irregular nuclear features (Sezary cells). MF typically presents with slowly progressing patch and plaque lesions. Detection of neoplastic CD4-positive T cells in peripheral blood (>1000 cells/microliter) is essential to establish a diagnosis of SS. Disease staging and assessment of therapy response in CTCL require a quantitative assessment of peripheral blood involvement in absolute number of neoplastic cells (Sezary cells) per microliter. Flow cytometry is now considered the method of choice to estimate the number of Sezary cells in peripheral blood, largely replacing the less reproducible and time-consuming morphologic quantitation of atypical lymphocytes on a peripheral blood smear, proposed by the International Society for Cutaneous Lymphomas, and the cutaneous lymphoma task force of the European Organization of Research and Treatment of Cancer. Typically, Sezary cells are immunophenotypically distinct and they are clonal.
Reference Values
An interpretive report will be provided. This test will be processed as a laboratory consultation. An interpretation of the immunophenotypic findings and, if available, morphologic features will be provided by a board-certified hematopathologist for every case.
Interpretation
An immunophenotypically distinct T-cell population is suggestive of clonality when the subset exhibits a restricted T-cell receptor beta-chain constant (TRBC) staining pattern defined as either 1) >85% of TRBC1-positive events, 2) <15% TRBC1-positive events, or 3) homogenous TRBC1-dim expression. The immunophenotype of the distinct T-cell population, its percentage of total lymphocytes, and its percentage of total analyzed events will be reported. The test will be resulted as "No phenotypically aberrant T-cell population detected" if there is no specific immunophenotype that allows the detection of TRBC-restricted T cells.
Cautions
Correlation with clinical features is necessary for diagnosis of Sezary syndrome. This analysis can only describe a cell population with aberrant phenotype, but the significance of this finding in isolation is uncertain.
Clinical Reference
1. Horna P, Deaver DM, Qin D, et al. Quantitative flow cytometric identification of aberrant T cell clusters in erythrodermic cutaneous T cell lymphoma. Implications for staging and prognosis. J Clin Pathol. 2014;67(5):431-436
2. Berg H, Otteson GE, Corley H, et al. Flow cytometric evaluation of TRBC1 expression in tissue specimens and body fluids is a novel and specific method for assessment of T-cell clonality and diagnosis of T-cell neoplasms. Cytometry B Clin Cytom. 2021;100(3):361-369
3. Horna P, Shi M, Olteanu H, Johansson U. Emerging role of T-cell receptor constant beta chain-1 (TRBC1) expression in the flow cytometric diagnosis of T-cell malignancies. Int J Mol Sci. 2021;22(4):1817
4. Wilcox RA. Cutaneous T-cell lymphoma: 2016 update on diagnosis, risk-stratification, and management. Am J Hematol. 2016;91(1):152-165. doi:10.1002/ajh.24233
5. Horna P, Olteanu H, Jevremovic D, et al. Single-antibody evaluation of T-cell receptor beta constant chain monotypia by flow cytometry facilitates the diagnosis of T-cell large granular lymphocytic leukemia. Am J Clin Pathol. 2021;156(1):139-148
6. Horna P, Shi M, Jevremovic D, Craig FE, Comfere NI, Olteanu H. Utility of TRBC1 expression in the diagnosis of peripheral blood involvement by cutaneous T-cell lymphoma. J Invest Dermatol. 2021;141(4):821-829
7. Scarisbrick JJ, Hodak E, Bagot M, et al. Blood classification and blood response criteria in mycosis fungoides and Sezary syndrome using flow cytometry: recommendations from the EORTC cutaneous lymphoma task force. Eur J Cancer. 2018;93:47-56
8. Illingworth A, Johansson U, Huang S, et al. International guidelines for the flow cytometric evaluation of peripheral blood for suspected Sézary syndrome or mycosis fungoides: Assay development/optimization, validation, and ongoing quality monitors. Cytometry B Clin Cytom. 2021;100(2):156-182
Method Description
Flow cytometry immunophenotyping of peripheral blood is performed using the following antibodies:
-Triage Panel: CD3, CD10, CD16, CD19, CD34, CD45, and kappa and lambda immunoglobulin light chains.
-Sezary Panel: CD2, CD3, CD4, CD5, CD7, CD8, CD26, CD45, and TRBC1.(Shi M, Jevremovic D, Otteson GE, Timm MM, Olteanu H, Horna P. Single antibody detection of T-cell receptor alpha-beta clonality by flow cytometry rapidly identifies mature T-cell neoplasms and monotypic small CD8-positive subsets of uncertain significance. Cytometry B Clin Cytom. 2020;98[1]:99-107)
-Possible additional panels performed per algorithmic approach.
Day(s) Performed
Monday through Saturday
Report Available
1 to 3 daysSpecimen Retention Time
14 daysPerforming Laboratory
Mayo Clinic Laboratories in Rochester
Test Classification
This test was developed using an analyte specific reagent. Its performance characteristics were determined by Mayo Clinic in a manner consistent with CLIA requirements. This test has not been cleared or approved by the US Food and Drug Administration.CPT Code Information
88184-Flow cytometry; first cell surface, cytoplasmic or nuclear marker x 1
88185-Flow cytometry; additional cell surface, cytoplasmic or nuclear marker (each)
88188-Flow Cytometry Interpretation, 9 to15 markers (if appropriate)
88189-Flow Cytometry Interpretation, 16 or more markers (if appropriate)
LOINC Code Information
| Test ID | Test Order Name | Order LOINC Value |
|---|---|---|
| SZDIA | Sezary Diagnostic Flow Cytometry, B | 101118-8 |
| Result ID | Test Result Name | Result LOINC Value |
|---|---|---|
| CK126 | Sezary Diagnostic | No LOINC Needed |
| CK127 | Final Diagnosis | 50398-7 |
| CK128 | Special Studies | 30954-2 |
| CK129 | Microscopic Description | 22635-7 |