Detecting a neoplastic clone in paraffin-embedded specimens associated with the common chromosome abnormalities seen in patients with B-cell lymphoblastic leukemia/lymphoma
Test Id | Reporting Name | Available Separately | Always Performed |
---|---|---|---|
_IL25 | Interphases, <25 | No, (Bill Only) | No |
_I099 | Interphases, 25-99 | No, (Bill Only) | No |
_I300 | Interphases, >=100 | No, (Bill Only) | No |
_PADD | Probe, +1 | No, (Bill Only) | No |
_PB02 | Probe, +2 | No, (Bill Only) | No |
_PB03 | Probe, +3 | No, (Bill Only) | No |
_PBCT | Probe, +2 | No, (Bill Only) | No |
This test includes a charge for application of the first probe set (2 fluorescence in situ hybridization [FISH] probes) and professional interpretation of results. Additional charges will be incurred for all reflex probes performed. Analysis charges will be incurred based on the number of cells analyzed per probe set. If no cells are available for analysis, no analysis charges will be incurred.
This FISH test allows different combinations of probes to be utilized based on the patient's age and clinical question, including the standard (diagnostic) B-cell lymphoblastic lymphoma (B-LBL) FISH panel and the individual B-LBL FISH probes (per client request).
The FISH panel for patients 30 years and younger includes testing for the following abnormalities using the FISH probes listed:
+9/9p-, CDKN2A/D9Z1
t(9;22) BCR/ABL1
11q23 rearrangement, MLL (KMT2A) break-apart
-17/17p-, TP53/D17Z1
t(1;19)(q23;p13), PBX1/TCF3
Hyperdiploidy, +4,+10,+17: D4Z1/D10Z1/D17Z1
t(12;21)(p13;q22), ETV6/RUNX1 fusion and iAMP21
14q32 rearrangement, IGH break-apart
8q24.1 rearrangement, MYC break-apart
If the initial FISH panel demonstrates normal or nonclassical abnormalities, the Philadelphia chromosome-like acute lymphoblastic leukemia (Ph-like ALL) panel will be performed.
The Ph-like ALL panel includes testing for the following kinase activating chromosome abnormalities, using the FISH probes listed below:
1q25 rearrangement, ABL2 break-apart
5q32 rearrangement, PDGFRB break-apart
9p24.1 rearrangement, JAK2 break-apart
9q34 rearrangement, ABL1 break-apart
The initial FISH panel for patients older than 30 years of age includes testing for the following abnormalities using the FISH probes listed:
t(9;22) BCR/ABL1
If BCR/ABL1 fusion is not observed, the Ph-like ALL panel will be performed. The Ph-like ALL panel includes testing for the following kinase activating chromosome abnormalities, using the FISH probes listed below:
1q25 rearrangement, ABL2 break-apart
5q32 rearrangement, PDGFRB break-apart
9p24.1 rearrangement, JAK2 break-apart
9q34 rearrangement, ABL1 break-apart
If the previous FISH probe sets demonstrate normal or nonclassical abnormalities, the following probe sets will be performed:
t(1;19)(q23;p13), PBX1/TCF3 fusion
Hyperdiploidy, +4,+10,+17: D4Z1/D10Z1/D17Z1
t(12;21)(p13;q22), ETV6/RUNX1 fusion and iAMP21
14q32 rearrangement, IGH break-apart
11q23 rearrangement, MLL (KMT2A) break-apart
When an MLL (KMT2A) rearrangement is identified, reflex testing will be performed to identify the translocation partner. Probes include identification of:
t(4;11)(q21;q23) AFF1/MLL
t(6;11)(q27;q23) MLLT4(AFDN)/MLL
t(9;11)(p22;q23) MLLT3/MLL
t(10;11)(p12;q23) MLLT10/MLL
t(11;19)(q23;p13.1) MLL/ELL
t(11;19)(q23;p13.3) MLL/MLLT1.
In the absence of BCR/ABL1 fusion, when an extra ABL1 signal is identified, reflex testing will be performed using the ABL1 break-apart probe set to evaluate for the presence or absence of an ABL1 rearrangement.
In the absence of ETV6/RUNX1 fusion, when an extra ETV6 signal is identified, reflex testing will be performed using the ETV6 break-apart probe set to evaluate for the presence or absence of an ETV6 rearrangement.
If a MYC rearrangement is identified, both the BCL2 and BCL6 probe sets will be performed.
For more information, see B-Lymphoblastic Leukemia/Lymphoma Algorithm.
Fluorescence In Situ Hybridization (FISH)
+4,+10,+17
17p- (17p deletion) or TP53
9p- (9p deletion) or CDKN2A or p16
ABL1 (9q34) rearrangement
ABL2 (1q25) rearrangement
BCR-ABL1 like ALL
Hyperdiploidy
Hypodiploid/pseudo-hyperdiploid
Hypotriploid/Near-Triploid
iAMP21
IGH (14q32) rearrangement
JAK2 (9p24.1) rearrangement
MLL or KMT2A (11q23) rearrangement
Ph-like ALL
Philadelphia-like ALL
t(1;19)(q23;p13.3) - PBX1/TCF3
t(10;11)(p12;q23) - MLLT10/MLL or AF10/MLL
t(11;19)(q23;p13.1) - MLL/ELL
t(11;19)(q23;p13.3) - MLL/MLLT1 or MLL/ENL
t(12;21)(p13;q22) - TEL/AML1 or ETV6/RUNX1
t(4;11)(q21;q23) - AFF1/MLL or AF4/MLL
t(6;11)(q27;q23) - MLLT4(AFDN)/MLL or AF6/MLL
t(9;11)(p22;q23) - MLLT3/MLL or AF9/MLL
t(9;22)(9q34;q11.2) - BCR/ABL1
MYC (8q24.1) rearrangement
12p13 rearrangement, ETV6
PDGFRB (5q23) rearrangement
This test includes a charge for application of the first probe set (2 fluorescence in situ hybridization [FISH] probes) and professional interpretation of results. Additional charges will be incurred for all reflex probes performed. Analysis charges will be incurred based on the number of cells analyzed per probe set. If no cells are available for analysis, no analysis charges will be incurred.
This FISH test allows different combinations of probes to be utilized based on the patient's age and clinical question, including the standard (diagnostic) B-cell lymphoblastic lymphoma (B-LBL) FISH panel and the individual B-LBL FISH probes (per client request).
The FISH panel for patients 30 years and younger includes testing for the following abnormalities using the FISH probes listed:
+9/9p-, CDKN2A/D9Z1
t(9;22) BCR/ABL1
11q23 rearrangement, MLL (KMT2A) break-apart
-17/17p-, TP53/D17Z1
t(1;19)(q23;p13), PBX1/TCF3
Hyperdiploidy, +4,+10,+17: D4Z1/D10Z1/D17Z1
t(12;21)(p13;q22), ETV6/RUNX1 fusion and iAMP21
14q32 rearrangement, IGH break-apart
8q24.1 rearrangement, MYC break-apart
If the initial FISH panel demonstrates normal or nonclassical abnormalities, the Philadelphia chromosome-like acute lymphoblastic leukemia (Ph-like ALL) panel will be performed.
The Ph-like ALL panel includes testing for the following kinase activating chromosome abnormalities, using the FISH probes listed below:
1q25 rearrangement, ABL2 break-apart
5q32 rearrangement, PDGFRB break-apart
9p24.1 rearrangement, JAK2 break-apart
9q34 rearrangement, ABL1 break-apart
The initial FISH panel for patients older than 30 years of age includes testing for the following abnormalities using the FISH probes listed:
t(9;22) BCR/ABL1
If BCR/ABL1 fusion is not observed, the Ph-like ALL panel will be performed. The Ph-like ALL panel includes testing for the following kinase activating chromosome abnormalities, using the FISH probes listed below:
1q25 rearrangement, ABL2 break-apart
5q32 rearrangement, PDGFRB break-apart
9p24.1 rearrangement, JAK2 break-apart
9q34 rearrangement, ABL1 break-apart
If the previous FISH probe sets demonstrate normal or nonclassical abnormalities, the following probe sets will be performed:
t(1;19)(q23;p13), PBX1/TCF3 fusion
Hyperdiploidy, +4,+10,+17: D4Z1/D10Z1/D17Z1
t(12;21)(p13;q22), ETV6/RUNX1 fusion and iAMP21
14q32 rearrangement, IGH break-apart
11q23 rearrangement, MLL (KMT2A) break-apart
When an MLL (KMT2A) rearrangement is identified, reflex testing will be performed to identify the translocation partner. Probes include identification of:
t(4;11)(q21;q23) AFF1/MLL
t(6;11)(q27;q23) MLLT4(AFDN)/MLL
t(9;11)(p22;q23) MLLT3/MLL
t(10;11)(p12;q23) MLLT10/MLL
t(11;19)(q23;p13.1) MLL/ELL
t(11;19)(q23;p13.3) MLL/MLLT1.
In the absence of BCR/ABL1 fusion, when an extra ABL1 signal is identified, reflex testing will be performed using the ABL1 break-apart probe set to evaluate for the presence or absence of an ABL1 rearrangement.
In the absence of ETV6/RUNX1 fusion, when an extra ETV6 signal is identified, reflex testing will be performed using the ETV6 break-apart probe set to evaluate for the presence or absence of an ETV6 rearrangement.
If a MYC rearrangement is identified, both the BCL2 and BCL6 probe sets will be performed.
For more information, see B-Lymphoblastic Leukemia/Lymphoma Algorithm.
Tissue
For testing non-paraffin bone marrow or blood samples from patients with B-cell acute lymphoblastic leukemia/lymphoma, order either BALPF / B-Cell Acute Lymphoblastic Leukemia/Lymphoma (ALL), FISH, Pediatric, Varies or BALAF / B-Cell Acute Lymphoblastic Leukemia/Lymphoma (ALL), FISH, Adult, Varies, depending on the patient's age. If non-paraffin bone marrow or blood sample is received for this test, it will be canceled, and either BALPF or BALAF, depending on patient's age, will be added and performed as the appropriate test.
For patients with B-cell lymphoma, order BLYM / B-Cell Lymphoma, FISH, Tissue.
Advise Express Mail or equivalent if not on courier service.
A reason for testing and pathology report are required for testing to be performed. Send information with specimen. Acceptable pathology reports include working drafts, preliminary pathology, or surgical pathology reports. The laboratory will not reject testing if this information is not provided; however. appropriate testing and interpretation may be compromised or delayed. If not provided, an appropriate indication for testing may be entered by Mayo Clinic Laboratories.
Question ID | Description | Answers |
---|---|---|
GC057 | Reason for Referral |
Submit only 1 of the following specimens:
Specimen Type: Tissue
Preferred: Tissue block
Collection Instructions: Submit a formalin-fixed, paraffin-embedded tumor tissue block. Blocks prepared with alternative fixation methods may be acceptable; 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: Tissue slides
Collection Instructions: 20 Consecutive, unstained, 5-micron thick sections placed on positively charged slides and 1 hematoxylin and eosin-stained slide.
If not ordering electronically, complete, print, and send 1 of the following forms with the specimen:
Fifteen consecutive, unstained, 5-micron thick sections placed on positively charged slides, and 1 hematoxylin and eosin-stained slide.
Specimen Type | Temperature | Time | Special Container |
---|---|---|---|
Tissue | Ambient (preferred) | ||
Refrigerated |
Detecting a neoplastic clone in paraffin-embedded specimens associated with the common chromosome abnormalities seen in patients with B-cell lymphoblastic leukemia/lymphoma
This test includes a charge for application of the first probe set (2 fluorescence in situ hybridization [FISH] probes) and professional interpretation of results. Additional charges will be incurred for all reflex probes performed. Analysis charges will be incurred based on the number of cells analyzed per probe set. If no cells are available for analysis, no analysis charges will be incurred.
This FISH test allows different combinations of probes to be utilized based on the patient's age and clinical question, including the standard (diagnostic) B-cell lymphoblastic lymphoma (B-LBL) FISH panel and the individual B-LBL FISH probes (per client request).
The FISH panel for patients 30 years and younger includes testing for the following abnormalities using the FISH probes listed:
+9/9p-, CDKN2A/D9Z1
t(9;22) BCR/ABL1
11q23 rearrangement, MLL (KMT2A) break-apart
-17/17p-, TP53/D17Z1
t(1;19)(q23;p13), PBX1/TCF3
Hyperdiploidy, +4,+10,+17: D4Z1/D10Z1/D17Z1
t(12;21)(p13;q22), ETV6/RUNX1 fusion and iAMP21
14q32 rearrangement, IGH break-apart
8q24.1 rearrangement, MYC break-apart
If the initial FISH panel demonstrates normal or nonclassical abnormalities, the Philadelphia chromosome-like acute lymphoblastic leukemia (Ph-like ALL) panel will be performed.
The Ph-like ALL panel includes testing for the following kinase activating chromosome abnormalities, using the FISH probes listed below:
1q25 rearrangement, ABL2 break-apart
5q32 rearrangement, PDGFRB break-apart
9p24.1 rearrangement, JAK2 break-apart
9q34 rearrangement, ABL1 break-apart
The initial FISH panel for patients older than 30 years of age includes testing for the following abnormalities using the FISH probes listed:
t(9;22) BCR/ABL1
If BCR/ABL1 fusion is not observed, the Ph-like ALL panel will be performed. The Ph-like ALL panel includes testing for the following kinase activating chromosome abnormalities, using the FISH probes listed below:
1q25 rearrangement, ABL2 break-apart
5q32 rearrangement, PDGFRB break-apart
9p24.1 rearrangement, JAK2 break-apart
9q34 rearrangement, ABL1 break-apart
If the previous FISH probe sets demonstrate normal or nonclassical abnormalities, the following probe sets will be performed:
t(1;19)(q23;p13), PBX1/TCF3 fusion
Hyperdiploidy, +4,+10,+17: D4Z1/D10Z1/D17Z1
t(12;21)(p13;q22), ETV6/RUNX1 fusion and iAMP21
14q32 rearrangement, IGH break-apart
11q23 rearrangement, MLL (KMT2A) break-apart
When an MLL (KMT2A) rearrangement is identified, reflex testing will be performed to identify the translocation partner. Probes include identification of:
t(4;11)(q21;q23) AFF1/MLL
t(6;11)(q27;q23) MLLT4(AFDN)/MLL
t(9;11)(p22;q23) MLLT3/MLL
t(10;11)(p12;q23) MLLT10/MLL
t(11;19)(q23;p13.1) MLL/ELL
t(11;19)(q23;p13.3) MLL/MLLT1.
In the absence of BCR/ABL1 fusion, when an extra ABL1 signal is identified, reflex testing will be performed using the ABL1 break-apart probe set to evaluate for the presence or absence of an ABL1 rearrangement.
In the absence of ETV6/RUNX1 fusion, when an extra ETV6 signal is identified, reflex testing will be performed using the ETV6 break-apart probe set to evaluate for the presence or absence of an ETV6 rearrangement.
If a MYC rearrangement is identified, both the BCL2 and BCL6 probe sets will be performed.
For more information, see B-Lymphoblastic Leukemia/Lymphoma Algorithm.
Specific cytogenetic abnormalities are identified in the majority of cases of B-ALL/LBL, either by conventional chromosome studies or fluorescence in situ hybridization studies. Each of the genetic subgroups is important to detect and can be critical prognostic markers. For example, a decision for early transplantation may be made if t(9;22)(q34;q11.2), KMT2A rearrangement, iAMP21, or a hypodiploid clone is identified. In contrast, if the ETV6/RUNX1 fusion or hyperdiploidy is identified, the patient has a more favorable prognosis and transplantation is rarely initially considered.
A newly recognized World Health Organization entity called BCR-ABL1-like ALL, also known as Philadelphia chromosome-like acute lymphoblastic leukemia, is increasing in importance due to the poor prognosis seen in pediatric, adolescent, and young adult ALL. Common features of this entity involve rearrangements with tyrosine kinase genes involving the following genes: ABL2, PDGFRB, JAK2, ABL1, CRLF2, and P2RY8, as well as deletions involving IKZF1. Patients who have failed conventional therapies have demonstrated favorable responses to targeted therapies when rearrangements involving these specific gene regions have been identified.
Evaluation of the MYC gene region is included in all diagnostic B-ALL panels to evaluate for Burkitt lymphoma. If a positive result is obtained, additional testing for the BCL2 and BCL6 gene regions will be performed.
Per National Comprehensive Cancer Network guidelines, a combination of cytogenetic and FISH testing is currently recommended in all pediatric and adult patients with B-ALL/LBL.
Table. Common Chromosome Abnormalities in B-cell Acute Lymphoblastic Leukemia/Lymphoma
Leukemia type | Cytogenetic change | Typical demographic | Risk category |
B-cell acute lymphoblastic leukemia | t(12;21)(p13;q22), ETV6/RUNX1 | Pediatric | Favorable |
Hyperdiploidy | Pediatric | Favorable | |
t(1;19)(q23;p13.3), PBX1/TCF3 | Pediatric | Intermediate to favorable | |
t(9;22)(q34;q11.2), BCR/ABL1 | All ages | Unfavorable | |
iAMP21, RUNX1 | Pediatric | Unfavorable | |
del(9p), CDKN2A | All ages | Unknown | |
t(11q23;var), MLL | All ages | Unfavorable | |
t(4;11)(q21;q23), AFF1/MLL | All ages | Unfavorable | |
t(6;11)(q27;q23), MLLT4(AFDN)/MLL | All ages | Unfavorable | |
t(9;11)(p22;q23), MLLT3/MLL | All ages | Unfavorable | |
t(10;11)(p12;q23), MLLT10/MLL | All ages | Unfavorable | |
t(11;19)(q23;p13.1), MLL/ELL | All ages | Unfavorable | |
t(11;19)(q23;p13.3), MLL/MLLT1 | All ages | Unfavorable | |
t(14q32;var), IGH | All ages | Variable | |
-17/17p-, TP53 | All ages | Unfavorable | |
t(8q24.1;var), MYC | Pediatric/ adolescent/ young adult | ||
Complex karyotype (> or =4 abnormalities) | Adult | Unfavorable | |
Low hypodiploidy/near triploidy | Adult | Unfavorable | |
Near-haploid/hypodiploid | All ages | Unfavorable | |
Philadelphia chromosome-like acute lymphoblastic leukemia (Ph-like ALL) | t(1q25;var), ABL2 | Pediatric/ adolescent/ young adult | Unfavorable |
t(5q32;var), PDGFRB | |||
t(9p24.1;var), JAK2 | |||
t(9q34;var), ABL1 |
An interpretive report will be provided.
A neoplastic clone is detected when the percent of cells with an abnormality exceeds the normal reference range for any given probe.
A positive result is not diagnostic for B-cell lymphoblastic lymphoma but may provide relevant prognostic information.
The absence of an abnormal clone does not rule out the presence of a neoplastic disorder.
Fixatives other than formalin (eg, Prefer, Bouin's) may not be successful for fluorescence in situ hybridization (FISH) assays. Although FISH testing will not be rejected due to non-formalin fixation, results may be compromised.
Paraffin-embedded tissues that have been decalcified may be unsuccessful for FISH analysis.
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 no FISH signals are observed post-hybridization, the case will be released indicating a lack of FISH results.
For each probe set, blinded fluorescence in situ hybridization analysis was performed on 20 to 25 normal paraffin-embedded, formalin-fixed tissue controls and between 2 and 20 paraffin-embedded, formalin-fixed tissue samples from patients diagnosed with B-cell lymphoblastic leukemia or lymphoma. Results from the 25 controls were used to generate the normal cutoff values.
1. Moorman AV, Harrison CJ, Buck GA, et al: Karyotype is an independent prognostic factor in adult acute lymphoblastic leukemia (ALL): analysis of cytogenetic data from patients treated on the Medical Research Council (MRC) UKALLXII/Eastern Cooperative Oncology Group (ECOG) 2993 trial. Blood 2007. Apr 15;109(8):3189-3197
2. Moorman AV: The clinical relevance of chromosomal and genetic abnormalities in B-cell precursor acute lymphoblastic leukemia. Blood Rev. 2012 May;26(3):123-135
3. Roberts KG, Li Y, Payne-Turner D, et al: Targetable kinase-activating lesions in Ph-like acute lymphoblastic leukemia. N Engl J Med. 2014 Sept 11;371(11):1005-1015
4. Mullighan CG: The genomic landscape of acute lymphoblastic leukemia in children and young adults. Hematology Am Soc Hematol Educ Program. 2014 Dec 5;2014(1):174-180
5. Arber DA, Orazi A, Hasserjian R, et al: The 2016 revision to the World Health Organization classification of myeloid neoplasms and acute leukemia. Blood. 2016 May 19;127(20):2391-2405
6. Swerdlow SH, Campo E, Harris NL, et al, eds: WHO Classification of Tumours. Vol 2.WHO Classification of Tumours of Haematopoietic and Lymphoid Tissues. 4th ed. IARC Press; 2017
This test is performed using commercially available and laboratory-developed probes. Deletion of the CDKN2A locus on chromosome 9, TP53 on chromosome 17, and gain of chromosomes 4, 10, and 17 are detected using enumeration strategy probes. Rearrangements involving ABL2, PDGFRB, MYC, JAK2, ABL1, MLL, ETV6, and IGH are detected using a dual-color break-apart (BAP) strategy probe. Dual-color, dual-fusion fluorescence in situ hybridization strategy probe sets are used to detect t(9;22), t(12;21), t(1;19), and in reflex testing when rearrangements of the MLL gene is detected. If a MYC gene region separation is identified, break-apart BCL2 and BCL6 will be evaluated using a dual-color BAP strategy probe.
Paraffin-embedded tissues 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 etcher on the back of the unstained slide to be assayed. For each probe set, the probes are hybridized to the appropriate target areas and 2 technologists each analyze 50 interphase nuclei (100 total) per probe set with the results expressed as the percent abnormal nuclei.(Unpublished Mayo method)
Monday through Friday
This test was developed, and its performance characteristics 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.
88271 x 2, 88291-DNA probe, each (first probe set), interpretation and report
88271 x 2-DNA probe, each; each additional probe set (if appropriate)
88271-DNA probe, each; coverage for sets containing 3 probes (if appropriate)
88271 x 2-DNA probe, each; coverage for sets containing 4 probes (if appropriate)
88271 x 3-DNA probe, each; coverage for sets containing 5 probes (if appropriate)
88274 w/modifier 52-Interphase in situ hybridization, <25 cells, each probe set (if appropriate)
88274-Interphase in situ hybridization, 25 to 99 cells, each probe set (if appropriate)
88275-Interphase in situ hybridization, 100 to 300 cells, each probe set (if appropriate)
Test Id | Test Order Name | Order LOINC Value |
---|---|---|
BLBLF | B-Lymphoblastic Leuk/Lymph, FISH,Ts | 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.
|
---|---|---|
609452 | Result Summary | 50397-9 |
609453 | Interpretation | 69965-2 |
609454 | Result Table | 93356-4 |
609455 | Result | 62356-1 |
GC057 | Reason for Referral | 42349-1 |
609456 | Specimen | 31208-2 |
609457 | Source | 31208-2 |
609458 | Tissue ID | 80398-1 |
609459 | Method | 85069-3 |
609460 | Additional Information | 48767-8 |
609461 | Disclaimer | 62364-5 |
609462 | Released By | 18771-6 |
Change Type | Effective Date |
---|---|
New Test | 2021-12-13 |