LEUKEMIA LYMPHOMA IMMUNOPHENOTYPING BY FLOW CYTOMETRY

Test Documentation
Components
The client is expected to define the panel or panels to be analyzed, using the Flow Cytometry Immunophenotypic Test Requisition form order (see appendix) and standardized panels (see below).
Correlative clinical (e.g. splenomegaly), laboratory information (e.g. lymphocytosis; monoclonal gammopathy) and the client's differential diagnosis are important aids to the interpretation of the flow cytometry results. When tissue diagnosis is known, it is helpful to include a copy of the surgical or cytology report. For bone marrow specimens, please include a copy of the peripheral blood findings. Please do not staple paperwork together.
Immunophenotyping Panels:
Please note, in the event of a markedly hypocellular sample, the original panel/order may have to be abbreviated meaning fewer tubes/antigens will be acquired in order to get a meaningful analysis. This will be done by the flow staff and pathologist.
Acute Leukemia (Lymphocytic or Mylogeneous Acute Leukemia):
Test Code: ACUTE
Includes: CD2, CD3, CD5, CD7, CD10, CD11b, CD13, CD14, CD15, CD16, CD19, CD20, CD33, CD34, CD45, CD56, CD61, CD117, CD235a and HLA-Dr.
Intracellular Markers: cyCD3, cyCD79a, cyMPO, cyTdT can be added.
Mature Lymphoid Leukemia/Lymphoma - Probably B Cell:
(Note: If BCELL is ordered, do not order SBCLL)
Test Code: BCELL
Includes: CD3, CD5, CD10, CD11c, CD19, CD20, CD23, CD38, CD45, Kappa & Lambda immunoglobulin light chains.
Hairy Cell Leukemia Panel:
(Note: If HAIRY is ordered, do not order BCELL)
Test Code: HAIRY
Includes: CD3, CD5, CD10, CD11c, CD19, CD20, CD22, CD23, CD25, CD38, CD45, Kappa & Lambda immunoglobulin light chains, CD103
Short B Cell Panel:
(Note: If SBCLL is ordered, do not order BCELL)
Test Code: SBCLL
Includes: CD5, CD19, CD23, CD45, Kappa & Lambda immunoglobulin light chains.
Mature Lymphoid Leukemia/Lymphoma Probably T:
Test Code: TCELL
Includes: CD2, CD3, CD4, CD5, CD7, CD8, CD25, CD26, CD45, CD56, CD (16+56), CD57
Plasma Cell Clonality (Multiple Myeloma):
Test Code: MYE
Includes: CD19, CD38, CD45, CD56, cytoplasmic Kappa and cytoplasmic-Lambda.
CSF Leukemia/Lymphoma Mature B Cell:
Test Code: SBCSF
Includes: CD5, CD19, CD23, CD45, Kappa & Lambda Light chain
Sezary Staging (Blood staging for Mycosis Fungoides/Sezary Cell):
Test Code: SEZ
Includes: CD3, CD4, CD7, CD8, CD26, CD45
Fluid T Subsets:
Test Code: FL348
Includes: CD3, CD4, CD8, CD45
Acute CSF:
Test Code: ACSFF
Includes: cyCD3, cyTdT, CD13, CD33, CD34, CD19, CD45

Specimen Required
General: While minimum & ideal amounts of sample, and transport conditions and times are suggested below, the flow lab will almost always attempt to perform the ordered flow analysis on minimal samples or markedly aged samples. However, in such instances, it may be impossible to render an interpretation due to loss of viability, loss of particular cell types and/or insufficient numbers of diagnostic cells that are analyzable by flow cytometry. Regardless of source please try to deliver sample within 24 hours of collection to Warde Laboratory even if the need for analysis is uncertain. The latter cases can be sent as "Flow Holds" (FLOWHOLD is Cerner code). Due to seasonal changes in temperature it is imperative that the proper temperature is maintained during shipping, because it directly affects cell viability. For example specimens requiring refrigeration should be shipped with a refrigerated "cold pack" or room temperature should be insulated in a way to prevent freezing during transit etc.
Please note: temperature stability may differ by specimen type.
Whole Blood: 1 Yellow ACD, 10 mL Solution A or 7 mL Solution B (1.0 mL minimum) room temperature.
Alt: EDTA, Heparin
Stability: ACD, EDTA, Heparin: Room Temperature: 72 hours; Refrigerate: 72 hours; Frozen: Unacceptable
Bone Marrow: 1 Green Top Heparin. 3.0 mL (1.0 mL minimum).
Alt: ACD, Marrow aspirate mixed 1:1 with Heparinized media (1000 u/mL Heparin, Mccoy's, or RPMI).
Lavender EDTA: Room Temperature: 48 hours; Refrigerate: 48 hours
Lymph Nodes, Tonsil, Spleen, Other Solid Tissues: 0.5 cm of viable solid tissue. Place sample into 15 mL conical tube filled with MEM, McCoys or RPMI. If media is not available, then place sample into saline soaked gauze and place into container with additional saline and ship immediately to lab.
Stability: Refrigerated: 24 hours
CSF: 1.0 mL CSF (0.5 mL minimum).
Stability: Refrigerate: 24 hours
Body Fluids: 15 mL fluid in sterile container (5 mL minimum).
Stability: Refrigerated: 24 hours

Alternate Specimen
EDTA, Heparin or ACD

Rejection Criteria
Frozen samples, cracked or leaking containers, sample >=72 hours from collection.

Methodology
Flow Cytometry

Reference Range
Interpretive report

Performed
Monday-Saturday

Turnaround Time2-3 days
Test CodeSee above
CPT-4 Code (s)
Acute Leukemia (Lymphocytic or Mylogeneous Acute Leukemia): 88184, 88185 x 19
Mature Lymphoid Leukemia/Lymphoma, Probably B Cell: 88184, 88185 x 10
Hairy Cell Leukemia Panel: 88184, 88185 x 14
Short B Cell Panel: 88184, 88185 x 5
Mature Lymphoid Leukemia/Lymphoma Probably T: 88184, 88185 x 10
Plasma Cell Clonality (Multiple Myeloma): 88184, 88185 x 5
CSF Leukemia/Lymphoma Mature B Cell: 88184, 88185 x 5
Sezary Staging: 88184, 88185 x 5
Fluid T Cell Subsets: 88184, 88185 x 3
CSF, Acute Leukemia, AML, B or T ALL: 88184, 88185 x 6
*plus 88185 for each additional cell marker added to primary panel.
LOINC Codes
Acute Leukemia   45270-6  
Comment/Interpretation   48767-8  
Diagnosis   34574-4  
Signing Pathologist   19139-5  

Warde
Medical
Laboratory