Test Id : AMLMF

This test includes a charge for the probe application, analysis, and professional interpretation of results for 1 probe set (2 individual fluorescence in situ hybridization [FISH] probes). Additional charges will be incurred for all reflex testing, if requested, or additional probe sets 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 test is performed using client-specified FISH probes and is not intended as a panel test. Specific probes must be listed in the probe request field. Reflex probes can be performed when appropriate if specified in the order request field.

When specified, any of the following probes will be performed:

inv(3) or t(3;3) or GATA2::MECOM fusion, request probe GATA2/MECOM

t(1;3)(p36;q21) or GATA2::PRDM16 fusion, request probe PRDM16/GATA2

t(3q26.2;var) or MECOM rearrangement, request probe MECOM break-apart

-5/5q-, request probe D5S630/EGR1

t(6;9)(p22.3;q34) or DEK::NUP214 fusion, request probe DEK/NUP214

-7/7q-, request probe D7Z1/D7S486

t(7;12)(q36;p13) or MNX1::ETV6 fusion, request probe MNX1/ETV6

t(8;16)(p11;p13) or KAT6A::CREBBP fusion, request probe KAT6A/CREBBP

t(8;21)(q21.3;q22) or RUNX1::RUNX1T1 fusion, request probe RUNX1T1/RUNX1

t(21q22;var) or RUNX1 rearrangement, request probe RUNX1 break-apart

t(9;22)(q34;q11) or BCR::ABL1 fusion, request probe ABL1/BCR

t(11p15;var) or NUP98 rearrangement, request probe NUP98 break-apart

t(11q23;var) or KMT2A rearrangement, request probe KMT2A break-apart

t(4;11)(q21;q23) or KMT2A:: AFF1 fusion, request probe AFF1/KMT2A

t(6;11)(q27;q23) or KMT2A::AFDN fusion, request probe AFDN/ KMT2A

t(9;11)(p21.3;q23) or KMT2A::MLLT3 fusion, request probe MLLT3/KMT2A

t(10;11)(p12;q23) or KMT2A::MLLT10 fusion, request probe MLLT10/KMT2A

t(11;19)(q23;p13.3) or KMT2A::MLLT1 fusion, request probe KMT2A/MLLT1

t(11;19)(q23;p13.1) or KMT2A::ELL fusion, request probe KMT2A/ELL

t(15;17)(q24;q21) or PML::RARA fusion, request probe PML/RARA

t(17q21;var) or RARA rearrangement, request probe RARA break-apart

inv(16) or t(16;16) or CBFB::MYH11 fusion, request probe MYH11/CBFB

t(16q22;var) or CBFB rearrangement, request probe CBFB break-apart

inv(16)(p13q24) or CBFA2T3::GLIS2 fusion, request probe CBFA2T3/GLIS2

-17/17p-, request probe TP53/D17Z1

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.

For more information see:

Acute Promyelocytic Leukemia: Guideline to Diagnosis and Follow-up

Acute Leukemias of Ambiguous Lineage Testing Algorithm

Acute Myeloid Leukemia: Testing Algorithm

This test is intended for instances when limited acute myeloid leukemia (AML) fluorescence in situ hybridization (FISH) probes are needed based on specific abnormalities or abnormalities identified in the diagnostic sample. The FISH probes to be analyzed must be specified on the ordering request. If targeted FISH probes are not included with this test order, test processing will be delayed and the test may be canceled and automatically reordered by the laboratory as either AMLFA / Adult Acute Myeloid Leukemia Panel, FISH, Varies or AMLFP / Pediatric Acute Myeloid Leukemia Panel, FISH, Varies depending on the age of the patient.

If only PML and RARA probes are requested or PML::RARA fusion is identified, the laboratory will automatically expedite testing. When only the PML/RARA probe set is ordered, the result will typically be reported the next business day.

Results will not be provided until testing is finalized. The laboratory is unable to provide preliminary results.

At diagnosis, conventional cytogenetic studies (CHRBM / Chromosome Analysis, Hematologic Disorders, Bone Marrow) and a complete AML FISH panel (either AMLFA or AMLFP) should be performed.

If a complete AML FISH panel is preferred for an adult patient, order AMLFA / Adult Acute Myeloid Leukemia Panel, FISH, Varies.

If a complete AML FISH panel is preferred for a pediatric patient, order AMLFP / Pediatric Acute Myeloid Leukemia Panel, FISH, Varies.

Minimal residual disease (MRD) monitoring in patients with AML known to have either t(15;17) with PML::RARA fusion, inv(16) or t(16;16) with CBFB::MYH11 fusion, t(8;21) with RUNX1::RUNX1T1 fusion, or t(9;22) with BCR::ABL1 fusion should be performed by quantitative reverse transcriptase polymerase chain reaction and NOT by FISH testing.

It is recommended that MRD monitoring in patients with AML be performed by AML-MRD flow cytometry rather than FISH testing using individual FISH probe sets. This is particularly true for the deletion/monosomy probe sets (5, 7, 17), which have cutoffs that exceed 10% of nuclei.

If this test is ordered and the laboratory is informed that the patient is age 30 years or younger AND is on a Children's Oncology Group protocol, this test will be canceled and automatically reordered by the laboratory as COGMF / Acute Myeloid Leukemia (AML), Children's Oncology Group Enrollment Testing, FISH, Varies.

For testing paraffin-embedded tissue samples from patients with AML/myeloid sarcoma, order MSTF / Myeloid Sarcoma, FISH, Tissue. If a paraffin-embedded tissue sample is submitted for this test, this test will be canceled and MSTF will be added and performed as the appropriate test.

At diagnosis, conventional cytogenetic studies (CHRBM / Chromosome Analysis, Hematologic Disorders, Bone Marrow) and a complete AMLFA / Adult Acute Myeloid Leukemia Panel, FISH, Varies or AMLFP / Pediatric Acute Myeloid Leukemia Panel, FISH, Varies should be performed, depending on patient's age. If there is limited specimen available, only fluorescence in situ hybridization testing will be performed.

Advise Express Mail or equivalent if not on courier service.

1. A list of targeted probes requested for analysis is required. Probes available for this test are listed in the Testing Algorithm section.

2. 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.

3. A flow cytometry and/or a bone marrow pathology report should be submitted with

each specimen. The laboratory will not reject testing if this information is not provided, but appropriate testing and interpretation may be compromised or delayed.

Submit only 1 of the following specimens:

Preferred

Specimen Type: Bone marrow

Container/Tube:

Preferred: Yellow top (ACD)

Acceptable: Green top (sodium heparin) or lavender top (EDTA)

Specimen Volume: 2 to 3 mL

Collection Instructions:

1. It is preferable to send the first aspirate from the bone marrow collection.

2. Invert several times to mix bone marrow.

3. Send bone marrow in original tube. Do not aliquot.

Acceptable

Specimen Type: Whole blood

Container/Tube:

Preferred: Yellow top (ACD)

Acceptable: Green top (sodium heparin) or lavender top (EDTA)

Specimen Volume: 6 mL

Collection Instructions:

1. Invert several times to mix blood.

2. Send whole blood in original tube. Do not aliquot.

• Acute Promyelocytic Leukemia: Guideline to Diagnosis and Follow-up
• Acute Leukemias of Ambiguous Lineage Testing Algorithm
• Acute Myeloid Leukemia: Testing Algorithm

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

Bone marrow: 1 mL; Whole blood: 2 mL

Detecting recurrent common chromosome abnormalities associated with acute myeloid leukemia (AML) using client-specified probe sets

As an adjunct to conventional chromosome studies in patients with AML

Evaluating specimens in which chromosome studies are unsuccessful

Identifying and tracking known chromosome abnormalities in patients with AML and monitoring response to therapy

This test includes a charge for the probe application, analysis, and professional interpretation of results for 1 probe set (2 individual fluorescence in situ hybridization [FISH] probes). Additional charges will be incurred for all reflex testing, if requested, or additional probe sets 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 test is performed using client-specified FISH probes and is not intended as a panel test. Specific probes must be listed in the probe request field. Reflex probes can be performed when appropriate if specified in the order request field.

When specified, any of the following probes will be performed:

inv(3) or t(3;3) or GATA2::MECOM fusion, request probe GATA2/MECOM

t(1;3)(p36;q21) or GATA2::PRDM16 fusion, request probe PRDM16/GATA2

t(3q26.2;var) or MECOM rearrangement, request probe MECOM break-apart

-5/5q-, request probe D5S630/EGR1

t(6;9)(p22.3;q34) or DEK::NUP214 fusion, request probe DEK/NUP214

-7/7q-, request probe D7Z1/D7S486

t(7;12)(q36;p13) or MNX1::ETV6 fusion, request probe MNX1/ETV6

t(8;16)(p11;p13) or KAT6A::CREBBP fusion, request probe KAT6A/CREBBP

t(8;21)(q21.3;q22) or RUNX1::RUNX1T1 fusion, request probe RUNX1T1/RUNX1

t(21q22;var) or RUNX1 rearrangement, request probe RUNX1 break-apart

t(9;22)(q34;q11) or BCR::ABL1 fusion, request probe ABL1/BCR

t(11p15;var) or NUP98 rearrangement, request probe NUP98 break-apart

t(11q23;var) or KMT2A rearrangement, request probe KMT2A break-apart

t(4;11)(q21;q23) or KMT2A:: AFF1 fusion, request probe AFF1/KMT2A

t(6;11)(q27;q23) or KMT2A::AFDN fusion, request probe AFDN/ KMT2A

t(9;11)(p21.3;q23) or KMT2A::MLLT3 fusion, request probe MLLT3/KMT2A

t(10;11)(p12;q23) or KMT2A::MLLT10 fusion, request probe MLLT10/KMT2A

t(11;19)(q23;p13.3) or KMT2A::MLLT1 fusion, request probe KMT2A/MLLT1

t(11;19)(q23;p13.1) or KMT2A::ELL fusion, request probe KMT2A/ELL

t(15;17)(q24;q21) or PML::RARA fusion, request probe PML/RARA

t(17q21;var) or RARA rearrangement, request probe RARA break-apart

inv(16) or t(16;16) or CBFB::MYH11 fusion, request probe MYH11/CBFB

t(16q22;var) or CBFB rearrangement, request probe CBFB break-apart

inv(16)(p13q24) or CBFA2T3::GLIS2 fusion, request probe CBFA2T3/GLIS2

-17/17p-, request probe TP53/D17Z1

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.

For more information see:

Acute Promyelocytic Leukemia: Guideline to Diagnosis and Follow-up

Acute Leukemias of Ambiguous Lineage Testing Algorithm

Acute Myeloid Leukemia: Testing Algorithm

Acute myeloid leukemia (AML) is one of the most common adult leukemias, with almost 10,000 new cases diagnosed per year. AML also comprises 15% of pediatric acute leukemia and accounts for the majority of infant (<1 year old) leukemia.

Several recurrent chromosomal abnormalities have been identified in AML with associated clinical significance. The most common chromosome abnormalities associated with AML include t(8;21), t(15;17), inv(16) or t(16;16), and abnormalities of the KMT2A gene at 11q23. The most common genes juxtaposed with KMT2A through translocation events in AML include AFDN- t(6;11), MLLT3- t(9;11), MLLT10- t(10;11), and ELL-t(11;19p13.1).

Acute myeloid leukemia can also evolve from myelodysplasia (MDS). Thus, the common chromosome abnormalities associated with MDS can also be identified in AML, which include inv(3) or t(3;3), -5/5q-, and -7/7q-. Overall, the recurrent chromosome abnormalities identified in patients with AML are observed in approximately 60% of diagnostic AML cases.

Conventional chromosome analysis is the gold standard for identification of the common, recurrent chromosome abnormalities in AML. However, some of the subtle rearrangements can be missed by karyotype, including inv(16) or t(16;16) and KMT2A rearrangements.

Fluorescence in situ hybridization analysis of nonproliferating (interphase) cells can be used to detect the common diagnostic and prognostic chromosome abnormalities observed in patients with AML.

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 set.

The absence of an abnormal clone does not rule out the presence of a neoplastic disorder.

This test is not approved by the US Food and Drug Administration, and it is best used as an adjunct to clinical and pathologic information.

Fluorescence in situ hybridization (FISH) is not a substitute for conventional chromosome studies because the latter detects chromosome abnormalities associated with other hematological disorders that would go undetected in a targeted acute myeloid leukemia FISH test.

Bone marrow is the preferred specimen type for this FISH test. If bone marrow is not available, a blood specimen may be used if there are circulating myeloblasts in the blood specimen (as verified by a hematopathologist).

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

1. Grimwade D, Hills RK, Moorman AV, et al. Refinement of cytogenetic classification in acute myeloid leukemia: determination of prognostic significance of rare recurring chromosomal abnormalities among 5876 younger adult patients treated in the United Kingdom Medical Research Council trials. Blood. 2010;116(3):354-365

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

3. Dohner H, Estey E, Grimwade D, et al. Diagnosis and management of AML in adults: 2017 ELN recommendations from an international expert panel. Blood. 2017;129(4):424-447. doi:10.1182/blood-2016-08-733196

4. Pollyea DA, Bixby D, Perl A, et al. NCCN Guidelines Insights: Acute Myeloid Leukemia, Version 2.2021. J Natl Compr Canc Netw. 2021;19(1):16-27. Published 2021 Jan 6. doi:10.6004/jnccn.2021.0002

This test is performed using commercially available and laboratory-developed fluorescence in situ hybridization (FISH) probes. Deletion or monosomy of chromosomes 5, 7, and 17 are detected using enumeration strategy probes. Rearrangements involving MECOM, ABL1, NUP98, KMT2A, CBFB, RARA, and RUNX1 are detected using dual-color break-apart (BAP) strategy probes. Dual-color FISH (D-FISH) strategy probe sets are used to detect t(1;3), inv(3) or t(3;3), t(6;9), t(7;12), t(8;16), t(8;21), t(9;22), t(15;17), inv(16) or t(16;16), and in reflex testing when rearrangements of the KMT2A gene are detected. For enumeration and BAP strategy probe sets, 100 interphase nuclei are scored; 200 interphase nuclei are scored when D-FISH probes are used. All results are expressed as the percent abnormal nuclei.(Unpublished Mayo method)

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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.

88271x2, 88275x1, 88291x1- FISH Probe, Analysis, Interpretation; 1 probe set

88271x2, 88275x1-FISH Probe, Analysis; each additional probe set (if appropriate)