LYMPHOMA/CHRONIC LYMPHOID LEUKEMIA PANEL, BY FLOW CYTOMETRY

Mnemonic

MISC

Synonym

ATTN FLOW - Flow Cytometry Studies - F LYMREQ - Acute Leukemia Panel - Acute Leukemia - Leukemia/Lymphoma Panel - L/L Panel - CLL Panel - B Cell Clonality - Lymphoma Protocol - R/O Lymphoma - Leukemia/Lymphoma for CLL - Chronic Lymphocytic Leukemia - Acute Leukemia - R/O CLL - Lymphoproliferative Disorder

Guidelines

This panel is designed to aid in the diagnosis and subcategorization of suspected or known leukemia, lymphoma or chronic lymphoid leukemia. 

Specimen Collection Requirements

Collect

Bone Marrow Aspirate: EDTA (lavender), Green (sodium heparin) or in RPMI. EDTA is preferred, but Green (sodium heparin is acceptable. RPMI is recommended if transit time of more than six hours is anticipated Also include 2 unstained aspirate smears.

Peripheral Blood: lavender (EDTA) top tube is preferred, but sodium heparin (green top) tube may be used.

Fresh Tissue: ~ cm3  ; Place in 5mL – 10mL RPMI.  Ensure that the tissue is completely immersed in the RPMI nutrient transport media.

Fine Needle Biopsy: 2-3 cores 1 cm long; Place in 5mL RPMI.  Ensure that the tissue is completely immersed in the RPMI nutrient transport media.

Fine Needle Aspirate: Place in 5mL RPMI in sterile container.

Fluids:  15.0-30.0 mL Body Fluid in sterile container. Do not send evac bottle.

CSF: 1-3 mL CSF in sterile container. Preferred: Centrifuge CSF, decant supernate and resuspend CSF cell pellet in 2 mL RPMI. Transport CSF cell pellet in RPMI at room temp ASAP.

BAL:  15.0 -30.0 mL BAL fluid in sterile container.

Handling

Maintain specimens at ambient temperature (20-25°C), avoid temperature extremes and do not centrifuge. Call Client Services ASAP at (503) 413-1234 or Toll Free (877) 270-5566 to arrange for PRIORITY pickup.

Specimen should be received within 24 hours of collection for optimal cell viability

Bone Marrow and Peripheral Blood: Always submit recent CBC with differential (collected within past 72 hours) and 2 unstained blood smears with request.

For complete collection and handling guidelines see Cytogenetics/Flow Cytometry/FISH Specimens for Leukemia, Lymphoma and Other Oncology Referrals

Preferred Volume

Bone Marrow: Collect 2-4 mL in sterile sodium heparinized syringe. Mix well by inverting and transfer into an EDTA tube (lavender top) or into RPMI for transport to the laboratory.

Peripheral Blood: 2-4mL EDTA (lavender top) anticoagulated blood.

Tissue: ~ 1 cm3 . Completely immerse fresh tissue in RPMI nutrient transport media.

Fine Needle Biopsy: 2-3 cores 1 – 2 cm long. Completely immerse fresh tissue in RPMI tissue culture medium.

Fine Needle Aspirate: Place in 5mL RPMI in sterile container. 

Fluids: 15.0-30.0 mL Body Fluid. Do not send evac bottle.

BAL: 15.0 mL BAL fluid in sterile container.

CSF: 1-3 mL CSF in sterile container.

Preferred: Centrifuge CSF, decant supernate and resuspend CSF cell pellet in 2 mL RPMI. Transport CSF cell pellet in RPMI at room temp ASAP.

Minimum Volume

Bone Marrow: Collect 1 mL in sterile sodium heparinized syringe. Mix well by inverting and transfer into an EDTA tube (lavender top) or into RPMI for transport to the laboratory.

Peripheral Blood: 2mL EDTA (lavender top) anticoagulated blood.

Tissue: ~ 1cm3 .  Completely immerse fresh tissue in RPMI tissue culture medium.

Fine Needle Biopsy: 1 core 1 – 2 cm long. Completely immerse fresh tissue in RPMI tissue culture medium.

Fluids: 5 mL Body Fluid. Do not send evac bottle.

Fine Needle Aspirate: Place in 5mL RPMI in sterile container. 

BAL: 5.0 mL BAL fluid in sterile container.

CSF: 1-3 mL CSF in sterile container.

Preferred: Centrifuge CSF, decant supernate and resuspend CSF cell pellet in 2 mL RPMI. Transport CSF cell pellet in RPMI at room temp ASAP.

Transport

Ambient (20-25°C)

Rejection Criteria

Bone Marrow: Clotted, severely hemolyzed or frozen specimens

Peripheral Blood: Clotted, severely hemolyzed or frozen, fixed specimens. Specimen in gel separator tubes.

Tissues, Fluids, Fine Needle Aspirates: Frozen, fixed or specimens in formalin.

CSF: Frozen or fixed specimens. Must have sufficient number of malignant cells for analysis

Specimen rejection: ONLY by Flow Cytometry personnel after consultation with the Hematopathologist.

Stability

Stability (collection to initiation of testing): Ambient (20°-25°C): 24 hours; Refrigerated: Acceptable 24 hours; Frozen: Unacceptable Fixed: Unacceptable

Testing Information

Note

Flow cytometric leukemia and lymphoma analysis may aid in identifying the tumor lineage for diagnostic and prognostic purposes. After review of the clinical history and morphology, a panel of markers is selected for each case by a board-certified pathologist. In most cases, the lineage can be identified as T-cell, B-cell, or myeloid and a diagnosis or differential diagnosis can be made. 

Available Markers*:

T-cell:
CD2, CD3, CD4, CD5, CD7, CD8, Cytoplasmic CD3
B-cell:
CD10, CD19, CD20, CD22, CD23, CD103, Kappa, Lambda, Cytoplasmic Kappa, Cytoplasmic Lambda
Myelo/Mono:
CD11b, CD13, CD14, CD15, CD33, CD64, CD117, myeloperoxidase (MPO)
Misc:
CD11c, CD16, CD25, CD34, CD36, CD38, CD41a, CD45, CD56, CD57, CD61, HLA-DR, TdT, CD138, CD45, CD79a.

*Not all markers will be reported in all cases.
Requests for specific markers to be run must be listed on manual requisition or by footnote for electronic orders. We do not offer individual marker identification separately outside of the markers in this panel.

A comprehensive flow cytometric Leukemia/Lymphoma assessment of tissue, fluid, or peripheral blood typically includes 14 or more markers; a bone marrow evaluation typically includes 15 or more markers. At the discretion of the pathologist, limited-cellularity samples (CSF, tissue/fluids) will be triaged and an appropriate number of markers will be run due to the irreplaceable nature of these samples. Limited-cellularity samples typically include a minimum number of markers (e.g., 8-15 markers).

The report will include a pathologist interpretation and a marker interpretation range corresponding to CPT codes of 2-8 markers, 9-15 markers, and 16+ markers interpreted. Charges apply per marker.

 

Remarks: A minimum of 10,000 viable cells is required for flow cytometry phenotyping of samples containing a very limited number of markers (may also be called antibodies or antigens). For low-count specimens, supplying clinical and diagnostic
information is especially important to help ensure that the most appropriate marker combinations are evaluated before the specimen is depleted of cells.


Bone marrow or whole blood: Provide specimen source, CBC, Wright stained smear (if available), clinical history, differential diagnosis, and any relevant pathology reports.


Tissue or fluid: Provide specimen source, clinical history, differential diagnosis, and any relevant pathology reports.


Follow up: If previous leukemia/lymphoma phenotyping was performed at another lab, the outside flow cytometry report and histograms (if possible) should accompany the specimen. 

 

Bone Marrow Requisitions and supplies including RPMI are available upon request.

Performed

 Monday-Friday (0800-1630)

Reported

Monday-Friday except holidays, 24-48 hours from receipt in Flow Lab

Method

Flow Cytometry

Reference Values

Reference Values

Interpretation provided.

CPT Codes

88184, 88185, 88189

Last Updated: Thursday, January 5, 2017 03:42:20 AM