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Test ID BLYM B-Cell Lymphoma, FISH, Tissue


Ordering Guidance


This test does not include a pathology consultation. If a pathology consultation is requested, order PATHC / Pathology Consultation, and appropriate testing will be added at the discretion of the pathologist and performed at an additional charge.

 

Mayo Hematopathology Consultants are involved in both the pre-analytic (tissue adequacy and probe selection, when applicable) and post-analytic (interpretation of fluorescence in situ hybridization [FISH] results in context of specific case, when applicable) phases.

 

This assay detects chromosome abnormalities observed in paraffin-embedded tissue samples of patients with B-cell lymphoma. If a non-paraffin embedded bone marrow or blood sample is received for this test, the test will be canceled, and BLPMF / B-Cell Lymphoma, Specified FISH, Varies will be added and performed as the appropriate test.

 

If either the break-apart MYC or the MYC/IGH D-FISH probe sets are requested in isolation, both probe sets will be performed concurrently to optimize the detection of MYC rearrangements.

 

For patients with T-cell lymphoma, order TLYM / T-Cell Lymphoma, FISH, Tissue.



Shipping Instructions


Advise Express Mail or equivalent if not on courier service.



Necessary Information


1. A pathology report is required for testing to be performed. If not provided, appropriate testing and/or interpretation may be compromised or delayed. Acceptable pathology reports include working drafts, preliminary pathology, or surgical pathology reports.

2. The following information must be included in the report provided.?

1. Patient name?

2. Block number - must be on all blocks, slides, and paperwork

3. Date of collection

4. Tissue Source

3. A reason for testing must be provided. If this information is not provided, an appropriate indication for testing may be entered by Mayo Clinic Laboratories.

A list of probes is required if select probes are necessary or if the patient is being tracked for known abnormalities. See Table in Clinical Information.



Specimen Required


Submit only 1 of the following specimens:

 

Preferred

Specimen Type: Tissue block

Collection Instructions: Submit a formalin-fixed, paraffin-embedded tumor tissue block. Blocks prepared with alternative fixation methods will be attempted but are less favorable for successful results; provide fixation method used.

Additional Information:

1. Paraffin-embedded specimens can be from any anatomic location (skin, soft tissue, lymph node, etc).

2. Bone specimens that have been decalcified will be attempted for testing, but the success rate is approximately 50%.

 

Acceptable

Specimen Type: Tissue slides

Slides: 1 Hematoxylin and eosin stained and 2 unstained?for each probe set

Collection Instructions:

1.  Include 1 hematoxylin and eosin-stained slide for the entire test order.

2. For each probe set ordered, submit 2 consecutive, unstained, 5 micron-thick sections placed on positively charged slides.

3. If ordering MYC, 4 unstained slides are necessary; the break-apart MYC and the MYC/IGH D-FISH are performed simultaneously.


Useful For

Detecting recurrent common chromosome abnormalities associated with various B-cell lymphomas in paraffin-embedded tissue specimens at diagnosis

Reflex Tests

Test ID Reporting Name Available Separately Always Performed
_PRAA Probe, Each Additional (BLYM) No, (Bill Only) No

Testing Algorithm

This test includes a charge for the probe application, analysis, and professional interpretation of results for one probe set (2 individual fluorescence in situ hybridization [FISH] probes). Additional charges will be incurred for all reflex or additional probe sets performed. No analysis charges will be incurred if an insufficient number of representative cells are available for analysis.

 

This FISH test allows different combinations of probes to be utilized based on the suspected lymphoma subtype, patient's age, and clinical question. The most appropriate probes to order are listed in the Common Chromosome Abnormalities in B-cell Lymphomas table in Clinical Information. Both the break apart MYC and the MYC/IGH dual-fusion FISH probes are analyzed simultaneously when MYC is requested. The BCL2/IGH FISH probe set will only be performed, at the laboratory's discretion, to resolve or confirm BCL2 rearrangement concerns.

 

Appropriate ancillary probes may be performed at consultant discretion to render comprehensive assessment. Any additional probes will have the results included within the final report and will be performed at an additional charge.

 

The following algorithms are available:

-Aggressive B-cell Lymphoma Diagnostic Algorithm

-Gastric MALT Lymphoma Diagnostic Algorithm

-Gastric MALT Posttherapy Follow-up Algorithm

Method Name

Fluorescence In Situ Hybridization (FISH)

Reporting Name

B-cell Lymphoma, FISH, Tissue

Specimen Type

Tissue

Specimen Minimum Volume

See Specimen Required

Specimen Stability Information

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

Reject Due To

All specimens will be evaluated at Mayo Clinic Laboratories for test suitability.

Clinical Information

Mature B-cell lymphoma can be low grade, intermediate grade, or high grade, and the prognosis and clinical course are highly variable. Genetic abnormalities have emerged as one of the most important prognostic markers in B-cell lymphomas and can aid in diagnosis. Several chromosome abnormalities and variants of these abnormalities have been associated with various lymphoma subtypes (see Table). Fluorescence in situ hybridization (FISH) permits the detection of recurrent gene rearrangements associated with various chromosome translocations and inversions in B-cell lymphoma. FISH is available for the specific B-cell lymphoma subtypes; see Table.

 

Table. Common Chromosome Abnormalities in B-cell Lymphomas

Lymphoma type

Chromosome abnormality

FISH probe

Burkitt (pediatric,

≤18 years old)

8q24.1 rearrangement

5'/3' MYC

t(2;8)(p12;q24.1)

IGK/MYC

t(8;14)(q24.1;q32)

MYC/IGH

t(8;22)(q24.1;q11.2)

MYC/IGL

3q27 rearrangement

3'/5' BCL6

18q21 rearrangement

3'/5' BCL2

Diffuse large B-cell, "double-hit"

8q24.1 rearrangement

5'/3' MYC

t(8;14)(q24.1;q32)

MYC/IGH

----Reflex: t(8;22)(q24.1;q11.2)

MYC/IGL

----Reflex: t(2;8)(p12;q24.1)

IGK/MYC

----Reflex: 3q27 rearrangement

3'/5' BCL6

----Reflex: 18q21 rearrangement

3'/5' BCL2

Large BCL IRF4 rearranged

6p24.3 rearrangement

5’/3' IRF4

18q21 rearrangement

3'/5' BCL2

3q27 rearrangement

3'/5' BCL6

Follicular

18q21 rearrangement

3'/5' BCL2

3q27 rearrangement

3'/5' BCL6

Predominantly diffuse subtype only: deletion of 1p36

TNFRSF14/1q22

Mantle cell

t(11;14)(q13;q32)

CCND1/IGH

11q13 rearrangement

5'/3' CCND1

Blastoid subtype only: deletion of 17p

TP53/D17Z1

Blastoid subtype only: 8q24.1 rearrangement

5'/3' MYC

Cyclin D1-negative subtype only: 12p13.32 rearrangement

5'/3' CCND2

MALT

18q21 rearrangement

5'/3' MALT1

Splenic marginal zone

Deletion of 7q

D7Z1/7q32

Deletion of 17p

TP53/D17Z1

Reference Values

An interpretive report will be provided.

Interpretation

A neoplastic clone is detected when the percent of cells with an abnormality exceeds the normal reference range for any given probe set.

 

Detection of an abnormal clone is supportive of a diagnosis of a B-cell lymphoma. The specific abnormality detected may help determine a B-cell lymphoma subtype and/or contribute to the prognosis.

 

The absence of an abnormal clone, or Negative result, does not rule out the presence of a neoplastic disorder or change the pathologic diagnosis.

Cautions

This test is not approved by the U.S. Food and Drug Administration and is best used as an adjunct to existing clinical and pathologic information.

 

Fixatives other than formalin (eg, Prefer, Bouin's) may not be successful for fluorescence in situ hybridization (FISH) assays. Non-formalin fixed specimens will not be rejected. 

 

Paraffin-embedded tissues that have been decalcified may not be successful for FISH analysis. The success rate of FISH studies on decalcified tissue is approximately 50%.

 

FISH studies will be attempted if sufficient tumor is present for analysis. The pathologist reviewing the hematoxylin and eosin-stained slide may find it necessary to cancel testing if insufficient tissue/tumor is available for testing.

 

If no FISH signals or a lack of sufficient tumor tissue are observed post-hybridization, the case will be released indicating a lack of FISH results.

Clinical Reference

Swerdlow SH, Campo E, Harris NL, eds, et al: WHO Classification of Tumours of Haematopoietic and Lymphoid Tissues. 4th ed. IARC; 2017. WHO Classification of Tumours. Vol 2

Method Description

This test is performed using either commercially available or laboratory-developed probes. Rearrangements involving MYC, BCL2, BCL6, CCND1, CCND2, IRF4, or MALT1 are detected using dual-color break-apart (BAP) strategy probes. Dual-color, dual-fusion fluorescence in situ hybridization strategy probe sets are used to detect t(11;14) and to evaluate the Immunoglobulin partners for the MYC gene.  Deletion of the 7q32 locus on chromosome 7, the TNFRSF14 locus on chromosome 1, and the TP53 locus on chromosome 17 are detected using enumeration strategy probes.

 

At the laboratory's discretion, the IGH/BCL2 probe will be performed when necessary to resolve or confirm BCL2 rearrangement concerns. IGH::BCL2 fusion is detected using a dual color, dual fusion probe set.

 

Paraffin-embedded tissue samples are cut at 5 microns and mounted on positively charged glass slides. The selection of tissue and the identification of target areas on the hematoxylin and eosin (H and E)-stained slide are performed by a pathologist. Using the H and E-stained slide as a reference, target areas are etched with a diamond-tipped engraving tool on the back of the unstained slide to be assayed. The probe set is hybridized to the appropriate target areas, and 2 technologists each independently analyze 50 interphase nuclei (100 total) with the results expressed as the percent of abnormal nuclei.(Unpublished Mayo method)

Day(s) Performed

Monday through Friday

Report Available

4 to 10 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.

CPT Code Information

88377 (if 1 probe set)

88377 x 2 (if 2 probe sets)

88377 x 3 (if 3 probe sets)

88377 x 4 (if 4 probe sets)

88377 x 5 (if 5 probe sets)

88377 x 6 (if 6 probe sets)

NY State Approved

Yes

Forms

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