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Test Code LLTOT Leukemia and Lymphoma Immunophenotyping, Technical Only, Tissue


Ordering Guidance


This test is available to clients through MayoAccess or MayoLink.

 

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



Shipping Instructions


Specimen must arrive within 4 days of collection.



Necessary Information


The following information is required:

1. Reason for testing

2. Tissue type

3. Tissue location

4. Surgical pathology case number



Specimen Required


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 ≤4 days/Refrigerated ≤4 days


Secondary ID

621505

Useful For

Evaluation of tissues for potential involvement by:

-Chronic lymphoproliferative disorders

-Malignant lymphomas

-Acute lymphoblastic leukemia

-Acute myelogenous leukemia

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 screening 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 tissue panel is initially performed to evaluate for monotypic B cells by kappa and lambda immunoglobulin light chain expression, CD5, CD10, CD19, CD20, and CD23.  Increased numbers of blasts and plasma cells are identified by CD45 expression along with side scatter gating.  The tissue panel also includes CD3, CD5, and CD7 antibodies to evaluate T cells. Additionally, viability is assessed on all tissue specimens using 7-AAD (7-Amino-Actinomycin D) exclusion.

 

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

 

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.

Method Name

Immunophenotyping

Reporting Name

Leukemia/Lymphoma; Tech Only Tissue

Specimen Type

Tissue

Specimen Minimum Volume

1 mm(3)

Specimen Stability Information

Specimen Type Temperature Time Special Container
Tissue Refrigerated (preferred)
  Ambient 

Reject Due To

Fixed, paraffin-embedded, or minced tissue Reject

Clinical Information

Cellular immunophenotyping, characterizing cells by using antibodies directed against cell surface markers, is generally regarded as a fundamental element in establishing a diagnosis of tissue involvement by hematolymphoid malignancies when used in conjunction with morphologic assessment. It is also an essential component in subclassification of hematolymphoid malignancies when present.

 

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

Reference Values

Not applicable

Interpretation

Report will include a summary of the procedure.

 

Normal tissues typically contain a mixture of B cells with polytypic surface immunoglobulin light chain expression and T cells with unremarkable expression of the T cell-associated antigens CD3, CD5, and CD7. Typically, no appreciable blast population is present by CD45 and side scatter analysis.

Cautions

It is well recognized that a negative flow cytometry result does not exclude tissue involvement by hematolymphoid malignancy. This may be attributable to sampling bias, although some malignancies, such as Hodgkin lymphoma, are not detected by this technique.

 

Viability will be assessed in all tissue specimens. Cases in which the viability is low (<50%) are prone to false-negative results and, therefore, must be interpreted with caution. In cases with viability less than 50%, testing will be attempted but may not be interpretable. Fine-needle aspiration and small biopsy specimens have a higher frequency of low cell counts and poor viability, which may be uninterpretable.

 

Even when abnormal, in most instances the results of flow cytometry are insufficient for complete subclassification of a hematolymphoid malignancy. Precise subclassification requires correlation with the histopathologic features in paraffin-embedded materials and, in some instances, the results of cytogenetic analyses.

 

The tissue used for flow cytometry cannot be subsequently submitted for histopathologic evaluation. For this reason, this technique should be avoided in small biopsy specimens.

Clinical Reference

1. Morice WG, Hodnefield JM, Kurtin PJ, Hanson CA. An unusual case of leukemic mantle cell lymphoma with a blastoid component showing loss of CD5 and aberrant expression of CD10. Am J Clin Pathol. 2004;122(1):122-127

2. Hanson CA. Acute leukemias and myelodysplastic syndromes. In: McClatchey KD, ed. Clinical Laboratory Medicine. Williams and Wilkins; 1994:939-969

3. Jaffe ES, Cossman J. Immunodiagnosis of lymphoid and mononuclear phagocytic neoplasms. In: Rose NR, Friedman H, Fahey JL, eds. Manual of Clinical Immunology. 3rd ed. ASM Press; 1987:779-790

4. Witzig TE, Banks PM, Stenson MJ, et al. Rapid immunotyping of B-cell non-Hodgkin's lymphomas by flow cytometry. A comparison with the standard frozen-section method. Am J Clin Pathol. 1990;94(3):280-286

5. Jevremovic D, Dronca RS, Morice WG, et al. CD5+ B-cell lymphoproliferative disorders: Beyond chronic lymphocytic leukemia and mantle cell lymphoma. Leuk Res. 2010;34(9):1235-1238

6. Jevremovic D, Olteanu H. Flow cytometry applications in the diagnosis of T/NK-cell lymphoproliferative disorders. Cytometry B Clin Cytom. 2019;96(2):99-115

7. Shi M, Jevremovic D, Otteson GE, Timm MM, Olteanu H, Horna P. Single antibody detection of T-Cell receptor alpha beta clonality by flow cytometry rapidly identifies mature T-Cell neoplasms and monotypic small CD8-positive subsets of uncertain significance. Cytometry B Clin Cytom. 2020;98(1):99-107

Method Description

Flow cytometric immunophenotyping of tissues is performed using the following antibodies:

Tissue Panel: CD3, CD5, CD7, CD10, CD19, CD20, CD23, CD45, 7-AAD, 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

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

-Acute Panel: CD2, CD3, CD5, CD7, CD13, CD15, CD19, CD20, CD33, CD34, CD45, CD56, 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

(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)

Day(s) Performed

Monday through Sunday

Report Available

1 to 4 days

Specimen Retention Time

Remaining tissue: 7 days

Performing Laboratory

Mayo Clinic Laboratories in Rochester

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.

CPT Code Information

88184-Flow cytometry; first cell surface, cytoplasmic or nuclear marker x 1

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
LLTOT Leukemia/Lymphoma; Tech Only Tissue 101119-6

 

Result ID Test Result Name Result LOINC Value
621506 LLTOT Result 69052-9
621507 Final Diagnosis 22637-3
621508 Special Studies 30954-2
621509 Microscopic Description 22635-7
CKR3 Reason for Referral 42349-1
CKS3 Tissue Type 31208-2
CKT3 Tissue Location 22633-2
CKPN3 Surgical Pathology Number 80398-1