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
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 |
AMLAB | Probe, Each Additional (AMLAF) | No, (Bill Only) | No |
AMLMB | Probe, Each Additional (AMLMF) | No, (Bill Only) | No |
AMLMF | AML, Specified FISH | Yes | No |
AMLAF | Adult AML, FISH | Yes | No |
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 |
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 to evaluate for monotypic B cells by kappa and lambda light chain expression, increased numbers of blast cells by CD34 and CD45 expression along with side scatter gating, and increased plasma cells by CD45 expression and 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 panel, 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 the 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 any of these tests. 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 as needed.
The following algorithms are available:
-Bone Marrow Staging for Known or Suspected Malignant Lymphoma Algorithm
-Acute Myeloid Leukemia: Testing Algorithm
-Acute Myeloid Leukemia: Relapsed with Previous Remission Testing Algorithm
-Acute Promyelocytic Leukemia: Guideline to Diagnosis and Follow-up
Immunophenotyping
Acute Leukemia -- Immunophenotyping, Flow Cytometry
B cell antigen
B cells, hematologic
B-cell clonality
Bone Marrow Consultation
Chronic Lymphocytic Leukemia, Immunophenotyping, Flow Cytometry
Flow Cytometry, Leukemia Immunophenotyping
Flow Cytometry, Lymphoma Immunophenotyping
Leukemia Immunophenotyping
Lymphocytoflow
Lymphoma Immunophenotyping by Flow Cytometry
Plasma cell clonality
T-cell clonality
T-cell receptor (TCR) flow cytometry
Anti-kappa
Anti-lambda
GLL Panel - Leukemia Immunophenotyping (ALWAYS order LCMS)
Granular Lymphocytic Leukemia (ALWAYS order LCMS)
KIR Panel - Leukemia Immunophenotyping (ALWAYS order LCMS)
LGL Panel - Leukemia Immunophenotyping (ALWAYS order LCMS)
NK Panel - Leukemia Immunophenotyping (ALWAYS order LCMS)
B-cell ALL minimal residual disease (MRD) detection
Mast Cell
Leukemia Lymphoma
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 to evaluate for monotypic B cells by kappa and lambda light chain expression, increased numbers of blast cells by CD34 and CD45 expression along with side scatter gating, and increased plasma cells by CD45 expression and 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 panel, 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 the 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 any of these tests. 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 as needed.
The following algorithms are available:
-Bone Marrow Staging for Known or Suspected Malignant Lymphoma Algorithm
-Acute Myeloid Leukemia: Testing Algorithm
-Acute Myeloid Leukemia: Relapsed with Previous Remission Testing Algorithm
-Acute Promyelocytic Leukemia: Guideline to Diagnosis and Follow-up
Varies
This test is appropriate for hematopoietic specimens only. For solid tissue specimens, order LLPT / Leukemia/Lymphoma Immunophenotyping, Flow Cytometry, Tissue.
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.
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.
Specimen must arrive within 96 hours of collection.
The following information is required:
1. Pertinent clinical history including reason for testing or clinical indication
2. Clinical or morphologic suspicion
3. Specimen source
4. Date and time of collection
5. For spinal fluid specimens: spinal fluid cell and differential counts are required.
Question ID | Description | Answers |
---|---|---|
CKR1 | Reason for Referral | |
CKS1 | Specimen Source |
Bone Marrow Peripheral blood Cerebral spinal fluid Pleural Peritoneal Pericardial Other body fluid |
Submit only 1 of the following specimens:
Specimen Type: Whole blood
Container/Tube:
Preferred: Yellow top (ACD solution A or B)
Acceptable: Green top (sodium heparin) or lavender top (EDTA)
Specimen Volume: 6 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 < or =96 hours
Specimen Type: Bone marrow
Container/Tube:
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 < or =96 hours
Specimen Type: Fluid
Sources: Serous effusions, pleural fluid, pericardial fluid, 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. 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 < or =96 hours
Specimen Type: Spinal fluid
Container/Tube: Sterile vial
Specimen Volume: 1 to 1.5 mL
Collection Instructions:
1. An original cytospin preparation (preferably unstained) must be included with the spinal fluid specimen so correlative morphologic evaluation can occur.
2. 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 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.
3. Label specimen as spinal fluid.
Specimen Stability Information: Refrigerated < or =96 hours
1. Hematopathology Patient Information (T676)
2. If not ordering electronically, complete, print, and send 1 of the following forms with the specimen:
-Hematopathology/Cytogenetics Test Request (T726)
-Benign Hematology Test Request (T755)
Blood: 3 mL
Bone Marrow, Spinal Fluid: 1 mL
Fluid from Serous Effusions: 5 mL
Gross hemolysis | Reject |
Specimen Type | Temperature | Time | Special Container |
---|---|---|---|
Varies | Varies |
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
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 to evaluate for monotypic B cells by kappa and lambda light chain expression, increased numbers of blast cells by CD34 and CD45 expression along with side scatter gating, and increased plasma cells by CD45 expression and 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 panel, 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 the 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 any of these tests. 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 as needed.
The following algorithms are available:
-Bone Marrow Staging for Known or Suspected Malignant Lymphoma Algorithm
-Acute Myeloid Leukemia: Testing Algorithm
-Acute Myeloid Leukemia: Relapsed with Previous Remission Testing Algorithm
-Acute Promyelocytic Leukemia: Guideline to Diagnosis and Follow-up
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.
An interpretive report will be provided.
This test will be processed as a laboratory consultation. An
Report will include a morphologic description, a summary of the procedure, the percent positivity of selected antigens, and an interpretive conclusion based on the correlation of the clinical history with the morphologic features and immunophenotypic results.
Specimens will be initially triaged to determine which, if any, of the immunophenotyping panels should be performed.
1. Jevremovic D, Dronca RS, Morice WG, et al: CD5+ B-cell lymphoproliferative disorders: Beyond chronic lymphocytic leukemia and mantle cell lymphoma. Leuk Res. 2010 Sep;34(9):1235-1238. doi: 10.1016/j.leukres.2010.03.020
2. Hanson CA: Acute leukemias and myelodysplastic syndromes. In: McClatchey KD, ed. Clinical Laboratory Medicine. Williams and Wilkins Inc; 1994:939-969
3. Jevremovic D, Olteanu H: Flow cytometry applications in the diagnosis of T/NK-cell lymphoproliferative disorders. Cytometry B Clin Cytom. 2019 Mar;96(2):99-115. doi: 10.1002/cyto.b.21768
4. Rosado FG, Morice WG, He R, Howard MT, Timm M, McPhail ED: Immunophenotypic features by multiparameter flow cytometry can help distinguish low grade B-cell lymphomas with plasmacytic differentiation from plasma cell proliferative disorders with an unrelated clonal B-cell process. Br J Haematol. 2015 May;169(3):368-376. doi: 10.1111/bjh.13303
5. Shi M, Ternus JA, Ketterling RP, et al: Immunophenotypic and laboratory features of t(11;14)(q13;q32)-positive plasma cell neoplasms. Leuk Lymphoma. 2018 Aug;59(8):1913-1919. doi: 10.1080/10428194.2017.1410885
6. Morice WG, Kimlinger T, Katzmann JA, et al: Flow cytometric assessment of TCR-Vbeta expression in the evaluation of peripheral blood involvement by T-cell lymphoproliferative disorders: a comparison with conventional T-cell immunophenotyping and molecular genetic techniques. Am J Clin Pathol. 2004 Mar;121(3):373-383. doi: 10.1309/3A32-DTVM-H640-M2QA
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 Jan;98(1):99-107. doi: 10.1002/cyto.b.21782
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 light chains
Possible Additional Panels:
-B-cell Panel: CD5, CD11c, CD19, CD20, CD22, CD23, CD38, CD45, CD103, CD200 and kappa and lambda 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, CD7, CD13, CD15, CD16, CD33, CD34, CD36, CD38, CD45, CD56, CD64, CD117, and HLA-DR
-B-cell ALL, minimal residual disease (MRD) panel: CD10, CD19, CD20, CD22, CD24, CD34, CD38, CD45, CD58, and CD66c
-Myeloperoxidase (MPO)/terminal deoxynucleotidyl transferase (TdT) (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 light chains
-Mast Cell Panel: CD2, CD25, CD69, CD117.(Keren D, McCoy JP, Carey J: Flow Cytometry in Clinical Diagnosis. 4th ed. American Society for Clinical Pathology; 2007; Betters DM: Use of flow cytometry in clinical practice. J Adv Pract Oncol. 2015 Sep-Oct;6[5]:435-440. doi: 10.6004/jadpro.2015.6.5.4)
Monday through Saturday
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.
88184-Flow cytometry; first cell surface, cytoplasmic or nuclear marker x 1
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)
Test Id | Test Order Name | Order LOINC Value |
---|---|---|
LCMS | Leukemia/Lymphoma, Phenotype | In Process |
Result Id | Test Result Name |
Result LOINC Value
Applies only to results expressed in units of measure originally reported by the performing laboratory. These values do not apply to results that are converted to other units of measure.
|
---|---|---|
18253 | Microscopic Description | 22635-7 |
18254 | Special Studies: | 30954-2 |
18255 | Final Diagnosis: | 34574-4 |
CK155 | LCMS Result | No LOINC Needed |
CKR1 | Reason for Referral | 42349-1 |
CKS1 | Specimen Source | 31208-2 |