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Test Code HLLFH Hematologic Disorders, Leukemia/Lymphoma; Flow Hold, Varies

Reporting Name

Heme Leukemia/Lymphoma; Flow Hold V

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
FCIMS Flow Cytometry Interp, 9-15 Markers No, (Bill Only) No
FCINS Flow Cytometry Interp,16 or greater No, (Bill Only) No
FCINT Flow Cytometry Interp, 2-8 Markers No, (Bill Only) No
AMLMF AML, Specified FISH Yes 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. Central time) 2 days after the collection date. For testing to be cancelled, the client must call 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, call 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).


In addition to reflexing flow cytometric panels, acute myeloid leukemia (AML) fluorescence in situ hybridization (FISH) testing for PML-RARA translocation t(15;17) may be added by the Mayo Clinic pathologist to exclude acute promyelocytic leukemia if there is morphologic suspicion or if blasts and promyelocytes are CD34-negative and HLA-DR-negative.


The triage panel is initially performed on peripheral blood, bone marrow, and fluid samples 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 panel can also evaluate T cells with CD3, CD5, and CD7. Additionally, viability is assessed on all tissue specimens using 7-AAD exclusion. 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.


These panels, together with the provided clinical history and morphologic review, are 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.


If no abnormalities are detected by the initial panel, no further flow cytometric assessment will be performed unless otherwise indicated by specific features of the clinical presentation or prior laboratory results.


Additional FISH or molecular testing may be recommended by the Mayo pathologist to facilitate diagnosis. The referring physician or pathologist will be contacted 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.

-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:

Confirmatory cytochemical stains are performed as needed.


The following algorithms are available:

-Bone Marrow Staging for Known or Suspected Malignant Lymphoma Algorithm

-Acute Promyelocytic Leukemia: Guideline to Diagnosis and Follow-up

Performing Laboratory

Mayo Clinic Laboratories in Rochester

Specimen Type


Ordering Guidance

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.


Bronchoalveolar lavage specimens submitted for evaluation for leukemia or lymphoma are appropriate to send 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.

Additional Testing Requirements

For bone marrow testing, if cytogenetic tests are desired along with this test request, an additional specimen should be submitted. It is important that the specimen be obtained, processed, and transported according to instructions for the other test.

Shipping Instructions

Specimen must arrive within 72 hours for fluids or 96 hours for peripheral blood, bone marrow, and tissue.

Necessary Information

The following information is required before the specimen will be processed:

a. Pertinent clinical history including reason for testing or clinical indication

b. A pathology/diagnostic report including the client surgical pathology case number.

c. Clinical or morphologic suspicion

d. Specimen source

e. Date and time of collection

f. For tissue specimens: tissue type and location are also required.

Specimen Required

Submit only 1 of the following specimens:


Specimen Type: Whole blood


Preferred: Yellow top (ACD solution A or B)

Acceptable: Green top (sodium heparin) or lavender top (EDTA)

Specimen Volume: 10 mL

Slides: If possible, include 5- to 10-unstained blood smears labeled with two unique identifiers

Collection Instructions:

1. Send whole blood specimen in original tube. Do not aliquot.

2. Label specimen as blood.

Specimen Stability Information: Ambient/Refrigerated <96 hours


Specimen Type: Bone marrow


Preferred: Yellow top (ACD solution A or B)

Acceptable: Green top (sodium heparin) or lavender top (EDTA)

Specimen Volume: 1 to 5 mL

Slides: If possible, include 5- to 10-unstained bone marrow aspirate smears labeled with two unique identifiers

Collection Instructions:

1. Submission of bilateral specimens is not required.

2. Label specimen as bone marrow.

Specimen Stability Information: Ambient/Refrigerated <96 hours


Specimen Type: Fluid

Sources: Serous effusions, pleural, pericardial, or abdominal (peritoneal fluid)

Note: Due to specimen stability, spinal fluid is not appropriate for this test.

Container/Tube: Body fluid container

Specimen Volume: 20 mL

Collection Instructions:

1. If possible, fluids 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

Supplies: Hank's Solution (T132)

Container/Tube: Sterile container with 15 mL of tissue culture medium (eg. Hank's balanced salt solution, 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

Specimen Minimum Volume

Blood: 3 mL
Bone Marrow: 1 mL
Fluid: 5 mL
Tissue: 1 mm(3) or larger biopsy

Specimen Stability Information

Specimen Type Temperature Time Special Container
Varies Varies

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.

Day(s) Performed

Monday through Saturday 

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

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)

LOINC Code Information

Test ID Test Order Name Order LOINC Value
HLLFH Heme Leukemia/Lymphoma; Flow Hold V In Process


Result ID Test Result Name Result LOINC Value
CK075 Final Diagnosis 34574-4
CK076 Special Studies 30954-2
CK077 Microscopic Description 22635-7
CK078 Flow Cytometry Testing No LOINC Needed

Report Available

2 to 4 days

Method Name


Additional Tests

Test ID Reporting Name Available Separately Always Performed
ADD1 Flow Cytometry, Cell Surface, Addl No, (Bill Only) Yes
FIRST Flow Cytometry, Cell Surface, First No, (Bill Only) Yes


1. Hematopathology Patient Information (T676)

2. If not ordering electronically, complete, print, and send a Hematopathology/Cytogenetics Test Request (T726) with the specimen.