Test ID LCMS Leukemia/Lymphoma Immunophenotyping by Flow Cytometry
Secondary ID
3287Useful For
Evaluating lymphocytoses of undetermined etiology
Identifying B- and T-cell lymphoproliferative disorders involving blood and bone marrow
Distinguishing acute lymphoblastic leukemia (ALL) from acute myeloid leukemia (AML)
Immunologic subtyping of ALL
Distinguishing reactive lymphocytes and lymphoid hyperplasia from malignant lymphoma
Distinguishing between malignant lymphoma and acute leukemia
Phenotypic subclassification of B- and T-cell chronic lymphoproliferative disorders, including chronic lymphocytic leukemia, mantle cell lymphoma, and hairy cell leukemia
Recognizing AML with minimal morphologic or cytochemical evidence of differentiation
Recognizing monoclonal plasma cells
Reflex Tests
Test ID | Reporting Name | Available Separately | Always Performed |
---|---|---|---|
VBETA | TCR V-BETA | No, (Bill Only) | No |
FIRST | Flow Cytometry, Cell Surface, First | No, (Bill Only) | No |
ADD1 | Flow Cytometry, Cell Surface, Addl | No, (Bill Only) | No |
FCINT | Flow Cytometry Interp, 2-8 Markers | No, (Bill Only) | No |
FCIMS | Flow Cytometry Interp, 9-15 Markers | No, (Bill Only) | No |
FCINS | Flow Cytometry Interp,16 or greater | No, (Bill Only) | No |
Testing Algorithm
When a blood, bone marrow, or fluid test is ordered, a screening panel and a professional interpretation will always be charged. The screening panel will be charged based on number of markers tested (FIRST for first marker, ADD1 for each additional marker). The interpretation will be based on markers tested in increments of 2 to 8, 9 to 15, or 16 and greater. 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 (ADD1) for each marker tested.
The triage panel is initially performed to evaluate for monotypic B cells by kappa and lambda light chain expression, increased numbers of blast cells by CD34 and CD45 expression along with side scatter gating, and increased plasma cells by CD45 expression and side scatter gating. The triage panel also includes antibodies to assess the number of CD3-positive T cells and CD16-positive/CD3-negative natural killer (NK) cells present. This triage panel also determines if there is an increase in the number of T cells that aberrantly coexpress CD16, an immunophenotypic feature of T-cell granular lymphocytic leukemia.
This panel, together with the provided clinical history and morphologic review, is used to determine what, if any, additional testing is needed for disease diagnosis or classification. If additional testing is required, it will be added per algorithm to fully characterize a disease state with a charge per unique antibody tested.
This test should not be ordered for bone marrow specimens being evaluated for possible involvement by a myelodysplastic syndrome (MDS) or a myelodysplastic/myeloproliferative neoplasm (MDS/MPN), including chronic myelomonocytic leukemia (CMML); order MYEFL / Myelodysplastic Syndrome by Flow Cytometry, Bone Marrow should be ordered.
In addition to reflexing flow cytometric panels, other studies may be added to facilitate diagnosis.
These include:
Cytogenetic FISH Studies:
-CCND1/IGH translocation t(11;14), to exclude mantle cell lymphoma in cases of CD5+CD23- B-cell lymphoproliferative disorder.
-PML-RARA translocation t(15;17), to exclude acute promyelocytic leukemia if there is morphologic suspicion and/or blasts and promyelocytes are CD34 and HLA-DR-negative.
-TCL-1 break-apart at 14q32, to exclude T-cell prolymphocytic leukemia in cases with CD4-positive T-cell lymphoproliferative disorder (phenotypic aberrancy or very tight CD4+ population with high CD4:CD8 ratio).
-MYC break-apart at 8q24, with or without IGH-BCL2 t(14;18) and BCL6 break-apart at 3q27, for suspected high grade B-cell lymphomas, based on morphologic assessment and immunophenotype (usually CD10-positive).
Molecular Genetic Studies:
-T-cell receptor gene rearrangement to examine clonality of T cells in cases showing phenotypically aberrant T-cell population.
Cytochemical Stains:
-Tartrate-resistant acid phosphatase (TRAP) stain to confirm hairy cell leukemia.
Note: FISH or molecular testing may be recommended by the Mayo pathologist in some cases. They will contact the referring physician or pathologist to confirm the addition of these tests.
The following algorithms are available in Special Instructions:
-Malignant Lymphoma, Guideline for Bone Marrow Staging Studies
-Acute Promyelocytic Leukemia: Guideline to Diagnosis and Follow-up
Special Instructions
Method Name
Immunophenotyping
Reporting Name
Leukemia/Lymphoma, PhenotypeSpecimen Type
VariesSpecimen must arrive within 48 hours of collection for spinal fluid, 96 hours for peripheral blood and bone marrow, or 72 hours for fluids.
This test is not appropriate for and cannot support diagnosis of sarcoidosis, hypersensitivity pneumonitis, interstitial lung diseases, or differentiating between pulmonary tuberculosis and sarcoidosis (requests for CD4/CD8 ratios). Specimens sent for these purposes will be rejected.
Bronchoalveolar lavage specimens submitted for evaluation for leukemia or lymphoma are appropriate to send for this test.
The following information is required:
1. Pertinent clinical history including reason for referral or clinical indication
2. Clinical or morphologic suspicion
3. Specimen source
4. Date and time of collection
Forms:
1. Hematopathology Patient Information Sheet (T676) in Special Instructions
2. If not ordering electronically, complete, print, and send 1 of the following forms with the specimen:
Hematopathology/Cytogenetics Test Request Form (T726) (http://www.mayomedicallaboratories.com/it-mmfiles/hematopathology-request-form.pdf)
Benign Hematology Test Request Form (T755) (http://www.mayomedicallaboratories.com/it-mmfiles/benign-hematology-test-request-form.pdf)
Submit only 1 of the following specimens:
Specimen Type: Blood
Container/Tube:
Preferred: Yellow top (ACD)
Acceptable: Heparin, EDTA
Specimen Volume: 10 mL
Collection Instructions:
1. Do not transfer blood to other containers.
2. Include 5 to 10 unstained blood smears, if possible.
3. Label specimen as blood.
Specimen Stability Information: Ambient <96 hours/Refrigerated ≤96 hours
Specimen Type: Bone marrow
Container/Tube:
Preferred: Yellow top (ACD)
Acceptable: ACD, Heparin, EDTA
Specimen Volume: 1-5 mL
Collection Instructions:
1. Submission of bilateral specimens is not required.
2. Include 5 to 10 unstained bone marrow aspirate smears, if possible.
3. Label specimen as bone marrow.
Specimen Stability Information: Ambient <96 hours/Refrigerated ≤96 hours
Additional Information: If cytogenetic tests are also desired when drawing LCMS / Leukemia/Lymphoma Immunophenotyping by Flow Cytometry, an additional specimen should be submitted. It is important that the specimen be obtained, processed, and transported according to instructions for the other required test.
Specimen Type: Fluid
Sources: Serous effusions
Container/Tube: Body fluid container
Specimen Volume: 20 mL
Collection Instructions:
1. If possible, the fluids other than spinal fluid should be anticoagulated with heparin (1 U/mL of fluid).
2. The volume of fluid necessary to phenotype the lymphocytes or blasts in serous effusions depends upon the cell count in the specimen. Usually 20 mL of pleural or peritoneal fluid is sufficient. Smaller volumes can be used if there is a high cell count.
3. Label specimen with fluid type.
Specimen Stability Information: Refrigerated <72 hours/Ambient ≤72 hours
Specimen Type: Spinal fluid
Container/Tube: Sterile vial
Specimen Volume: 1-1.5 mL
Collection Instructions:
1. An original cytospin preparation (preferably unstained) must be included with the spinal fluid specimen so correlative morphologic evaluation can occur.
2. The volume of fluid necessary to phenotype the lymphocytes or blasts in spinal fluid depends upon the cell count in the specimen. A cell count should be determined and submitted with the specimen. Usually 1 to 1.5 mL of spinal fluid is sufficient. Smaller volumes can be used if there is a high cell count. If cell count is <10 cells/mcL, a larger volume of spinal fluid may be required. When cell counts drop below 5 cells/mcL, the immunophenotypic analysis may not be successful.
3. Label specimen as spinal fluid.
Specimen Stability Information: Refrigerated <48 hours/Ambient ≤48 hours
Additional Information: Spinal fluid cell and differential counts are required.
Specimen Minimum Volume
Blood: 3 mL; Bone Marrow, Spinal Fluid: 1 mL; Fluid from Serous Effusions: 5 mL
Specimen Stability Information
Specimen Type | Temperature | Time |
---|---|---|
Varies | Varies |
Clinical Information
Diagnostic hematopathology has become an increasingly complex subspecialty, particularly with neoplastic disorders of blood and bone marrow. While morphologic assessment of blood smears, bone marrow smears, and tissue sections remains the cornerstone of lymphoma and leukemia diagnosis and classification, immunophenotyping is a very valuable and important complementary tool.
Immunophenotyping hematopoietic specimens can help resolve many differential diagnostic problems posed by the clinical or morphologic features.
This test is appropriate for hematopoietic specimens only. If your specimen is a solid tissue, order LLPT / Leukemia/Lymphoma Immunophenotyping by Flow Cytometry, Tissue.
Reference Values
An interpretive report will be provided.
This test will be processed as a laboratory consultation. An interpretation of the immunophenotypic findings and correlation with the morphologic features will be provided by a hematopathologist for every case.
Cautions
Specimens will be initially triaged to determine which, if any, of the immunophenotyping panels should be performed.
Day(s) Performed
Specimens are processed and reported Monday through Saturday
Report Available
1 dayPerforming Laboratory

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 U.S. 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)
88187-Flow Cytometry Interpretation, 2 to 8 Markers (if appropriate)
88188-Flow Cytometry Interpretation, 9 to 15 Markers (if appropriate)
88189-Flow Cytometry Interpretation, 16 or More Markers (if appropriate)