Test ID HLLFH Hematologic Disorders, Leukemia/Lymphoma; Flow Hold Varies
Useful 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 |
---|---|---|---|
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
This test is designed to delay the start of leukemia/lymphoma immunophenotyping until the preliminary assessment is completed. Specimens are held in the laboratory until noon (12 p.m. CST) 2 days after the collection date. For testing to be cancelled, the client must call Mayo Medical Laboratories at 800-533-1710. The testing process will be initiated and fully charged if no notification is received within this time period. To expedite the beginning of testing, please call Mayo Medical Laboratories at 800-533-1710.
The testing process begins with a screening panel. The screening panel will be charged based on the 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 for each marker tested (FIRST if applicable, ADD1 if applicable).
The triage panel is initially performed to evaluate for monotypic B cells by kappa and lambda light chain expression, increased numbers of blasts 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, further 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.
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) should be ordered as MYEFL / Myelodysplastic Syndrome by Flow Cytometry, Bone Marrow, not this test.
In addition to reflexing flow cytometric panels, FISH or molecular testing may be recommended by the Mayo pathologist to facilitate diagnosis. They will contact the referring physician or pathologist to confirm the addition of these tests.
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:
-TRAP stain to confirm hairy cell leukemia.
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
Heme Leukemia/Lymphoma; Flow Hold VSpecimen Type
VariesSpecimen must arrive within 96 hours for peripheral blood, bone marrow, and tissue, or 72 hours for fluids.
Due to specimen stability, spinal fluid is not appropriate for this test.
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 a Hematopathology/Cytogenetics Test Request Form (T726) with the specimen
(http://www.mayomedicallaboratories.com/it-mmfiles/hematopathology-request-form.pdf)
Submit only 1 of the following specimens:
Specimen Type: Blood
Container/Tube:
Preferred: Yellow top (ACD solution B)
Acceptable: ACD (solution A), 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/Refrigerated <96 hours
Specimen Type: Bone marrow
Container/Tube:
Preferred: Yellow top (ACD solution B)
Acceptable: ACD (solution A), 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/Refrigerated <96 hours
Additional Information: If cytogenetic tests are also desired when drawing HLLFH / Hematologic Disorders, Leukemia/Lymphoma; Flow Hold Varies, 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/Ambient <72 hours
Specimen Type: Tissue
Container/Tube: Sterile container with 15 mL of tissue culture medium (eg, Hank's balanced salt solution [T132], RPMI, or equivalent)
Specimen Volume: 5 mm(3) or larger biopsy
Collection Instructions:
1. Send intact specimen (do not mince).
2. Specimen cannot be fixed.
Specimen Stability Information: Ambient/Refrigerated <96 hours
Additional Information:
1. Date, time of collection, tissue type, and location are required.
2. A pathology/diagnostic report including the client surgical pathology case number, a brief history, reason for referral or clinical suspicion are required before the specimen will be processed.
Specimen Minimum Volume
Blood: 3 mL; Bone Marrow: 1 mL; Fluid from Serous Effusions: 5 mL; Tissue: 5 mm(3) or larger biopsy
Specimen Stability Information
Specimen Type | Temperature | Time |
---|---|---|
Varies | Varies |
Clinical Information
Immunophenotyping hematopoietic specimens can help resolve many differential diagnostic problems posed by the clinical or morphologic features.
Reference Values
When performed, 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.
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
2 daysPerforming 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
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)