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Test Code LLTOF Leukemia and Lymphoma Phenotyping, Technical Only, Varies

Reporting Name

Leukemia/Lymphoma; Tech Only Flow

Useful For

Evaluating lymphocytoses of undetermined etiology

 

Identifying B- and T-cell lymphoproliferative disorders involving blood and bone marrow

 

Distinguishing acute lymphoblastic leukemia from acute myeloid leukemia (AML)

 

Immunologic subtyping of acute leukemias

 

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

 

This test is not intended for detection of minimal residual disease below 5% blasts.

Additional Tests

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

Reflex Tests

Test ID Reporting Name Available Separately Always Performed
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

Note: This test is only available to clients who have MayoAccess or MayoLink.

 

The client is responsible for the interpretation and billing of the professional component; Mayo Clinic will bill the technical component only.

 

The testing process begins with a screening panel. The panel will be charged based on the number of markers tested (FIRST for first marker, ADD1 for each additional marker). Additional testing may be performed at an additional charge for each marker tested (ADD1, as applicable) if needed to fully characterize a disease state or clarify any abnormalities from the screening panel.

 

The triage panel is initially performed to evaluate for monotypic B cells by kappa and lambda immunoglobulin light chain expression, increased numbers of blasts by CD34 and CD45 expression along with side scatter gating, and increased plasma cells by CD45 expression with 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 initial testing, 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.

 

Cases requiring the testing for granular lymphocytic leukemia (killer-cell immunoglobulin-like receptor panel) will have an interpretation added and performed by a Mayo Clinic pathologist at an additional charge.

 

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

 

The following algorithms are available:

-Bone Marrow Staging for Known or Suspected Malignant Lymphoma Algorithm

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

Method Name

Immunophenotyping

Performing Laboratory

Mayo Clinic Laboratories in Rochester

Specimen Type

Varies


Ordering Guidance


This test is available to clients through MayoAccess or MayoLink.

 

For B-cell acute lymphoblastic leukemia minimal residual disease testing in either blood or bone marrow, order BALLM / B-Cell Lymphoblastic Leukemia Monitoring, Minimal Residual Disease Detection, Flow Cytometry, Varies.

 

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.

 

This test is not intended for product of conception (POC) specimens. For POC specimens see CMAPC / Chromosomal Microarray, Autopsy, Products of Conception, or Stillbirth.



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 4 days of collection.



Necessary Information


The following information is required:

1. Reason for testing

2. Specimen source

3. For spinal fluid specimens: spinal fluid cell and differential counts are required



Specimen Required


Submit only 1 of the following specimens:

 

Specimen Type: Whole blood

Container/Tube:

Preferred: Yellow top (ACD solution A or B)

Acceptable: Lavender top (EDTA) or green top (sodium heparin)

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 ≤4 days/Refrigerated ≤4 days

 

Specimen Type: Bone marrow

Container/Tube:

Preferred: Yellow top (ACD solution A or B)

Acceptable: Lavender top (EDTA) or green top (sodium heparin)

Specimen Volume: 1 to 5 mL

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

Collection Instructions:

1. Submission of bilateral specimens is not required.

2. Send bone marrow specimen in original tube. Do not aliquot.

3. Label specimen as bone marrow.

Specimen Stability Information: Ambient ≤4 days/Refrigerated ≤ 4 days

 

Specimen Type: Fluid

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

Container/Tube: Body fluid container

Specimen Volume: 20 mL

Collection Instructions:

1. If possible, fluids other than spinal fluid should be anticoagulated with heparin (1 U/mL of fluid).

2. Label specimen with fluid type.

Additional Information: 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.

Specimen Stability Information: Refrigerated ≤4 days/Ambient ≤4 days

 

Specimen Type: Spinal fluid

Container/Tube: Sterile vial

Specimen Volume: 1 to 1.5 mL

Collection Instructions:

1. An original cytospin preparation (preferably unstained) should be included with the spinal fluid specimen so correlative morphologic evaluation can occur.

2. Label specimen as spinal fluid.

Specimen Stability Information: Refrigerated /Ambient ≤4 days

Additional Information: 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 the cell count is less than 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.


Specimen Minimum Volume

Blood: 3 mL
Bone marrow: 0.5 mL
Spinal fluid: 1 mL
Fluid from serous effusions: 5 mL

Specimen Stability Information

Specimen Type Temperature Time Special Container
Varies Varies

Reject Due To

Gross hemolysis Reject
Fully clotted whole blood Reject

Reference Values

Not applicable

Day(s) Performed

Monday through Sunday

CPT Code Information

88184-Flow cytometry; first cell surface, cytoplasmic or nuclear marker

88185-Flow cytometry; additional cell surface, cytoplasmic or nuclear marker (each)

Additional CPTs may be added if consultative help is needed with the case, or algorithm dictates Mayo consultant involvement.

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
LLTOF Leukemia/Lymphoma; Tech Only Flow 101119-6

 

Result ID Test Result Name Result LOINC Value
CK071 Flow Cytometry 69052-9
CK072 Final Diagnosis 22637-3
CK073 Microscopic Description 22635-7
CK074 Special Studies 30954-2
CKR2 Reason for Referral 42349-1
CKS2 Specimen Source 31208-2

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.

 

This is a technical only test and does not include interpretation unless reflex testing is performed. At any point, clients may request to have a Mayo Clinic hematopathologist provide an interpretation at an additional charge.

Interpretation

Report will include a summary of the procedure.

Cautions

Specimens will be initially screened to determine which, if any, of the immunophenotyping panels should be performed.

Method Description

Flow cytometric immunophenotyping of peripheral blood, bone marrow, and body fluids is performed using the following antibodies:

 

Triage Panel: CD3, CD10, CD16, CD19, CD34, CD45, and kappa and lambda immunoglobulin light chains.

 

Possible Additional Panels: Performed per algorithmic approach

-B-cell Panel: CD5, CD11c, CD19, CD20, CD22, CD23, CD38, CD45, CD103, CD200 and kappa and lambda immunoglobulin light chains

-T-cell Panel: CD2, CD3, CD4, CD5, CD7, CD8, CD45, TRBC1, and gamma/delta

-Sezary Panel: CD2, CD3, CD4, CD5, CD7, CD8, CD26, CD45, and TRBC1.

-Killer-cell immunoglobulin-like receptor (KIR) Panel: CD3, CD8, CD16, CD56, CD57, CD94, CD158a, CD158b, CD158e (p70), and NKG2a

-Acute Panel: CD2, CD7, CD13, CD15, CD16, CD33, CD34, CD36, CD38, CD45, CD56, CD64, CD117, and HLA-DR

-B-cell acute lymphocytic leukemia (ALL) Panel: CD10, CD19, CD20, CD22, CD24, CD34, CD38, CD45, CD58, and CD66c

-Myeloperoxidase/terminal deoxynucleotidyl transferase (MPO/TdT) Panel: cytoplasmic CD3, CD13, cytoplasmic CD22, CD34, CD45, cytoplasmic CD79a, nuclear TdT, and cytoplasmic MPO

-Plasma Cell Panel: CD19, CD38, CD45, CD138, and cytoplasmic kappa and lambda immunoglobulin light chains

-Mast Cell Panel (bone marrow only): CD2, CD25, CD69, CD117.(Keren P, McCoy JP, Carey J, eds. Flow Cytometry in Clinical Diagnosis. 4th ed. ASCP Press; 2007; Betters DM. Use of flow cytometry in clinical practice. J Adv Pract Oncol. 2015;6[5]:435–440)

Report Available

1 to 4 days

Specimen Retention Time

Remaining blood/bone marrow: 14 days; Remaining fluid: 7 days

Test Classification

This test was developed and its performance characteristics determined by Mayo Clinic in a manner consistent with CLIA requirements. It has not been cleared or approved by the US Food and Drug Administration.

Forms

1. Hematopathology Patient Information (T676)

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