Test ID: FAML
Acute Myeloid Leukemia (AML), FISH

Detecting a neoplastic clone associated with the common chromosome anomalies seen in patients with acute myeloid leukemia or other myeloid malignancies

Evaluating specimens in which standard cytogenetic analysis is unsuccessful

Identifying and tracking known chromosome anomalies in patients with myeloid malignancies and tracking response to therapy

See Acute Promyelocytic Leukemia: Guideline to Diagnosis and Follow-up in Special Instructions.

When this test is ordered, a charge for 2 FISH probes and interpretation is included. If additional probes or the entire panel are ordered, additional probe charges will be added.

Please indicate if the entire panel is to be performed. This is suggested for patients with a new diagnosis of acute myeloid leukemia. If the patient is being treated for known anomalies, indicate which anomalies need to be investigated.

Indicate subtype as well as which anomalies need to be investigated from the following profile:

t(8:21), [M2], RUNX1T1/RUNX1

t(15:17), [M3], PML/RARA

t(11q23:var), [M0-M7], MLL

inv(16), [M4, Eos], MYH11/CBFB

+8, [M0-M7], Cen8/MYC

t(6;9), [M2,M4], DEK/NUP214

inv(3), [M1,2,4,6,7], RPN1/MECOM

t(8;16), [M4,M5], MYST3/CREBBP

t(1;22), [M7], RBM15/MKL1*

-5/5q-, D5S630/EGR1

-7/7q-, D7S486/Cen7

13q-, D13S319/LAMP1

17p-, TP53/Cen17

20q-, D20S108/20qter

t(9;22), BCR/ABL1

*The RBM15/MKL1 probe set will only be used to test patients with a suspected or confirmed diagnosis of M7 or to confirm a t(1;22) identified by chromosome analysis.

-When an MLL rearrangement is identified, reflex testing is performed to identify the translocation partner. Probes include identification of t(6;11)(q27;q23) MLLT4/MLL, t(9;11)(p22;q23) MLLT3/MLL, t(10;11)(p13;q23) MLLT10/MLL, t(11;16)(q23;p13.3) MLL/CREBBP, t(11;19)(q23;p13.1) MLL/ELL, or t(11;19)(q23;p13.3) MLL/MLLT1.

-When 3 copies of MECOM are observed with no fusion with RPN1 reflex testing using the MECOM/RUNX1 probe set is performed to identify a potential t(3;21)(q26.2;q22) rearrangement.

-When 3 copies of RPN1 are observed with no fusion with MECOM, reflex testing using the PRDM16/RPN1 probe set is performed to identify a potential t(1;3)(p36;q21).

• Cytogenetics Hematologic FISH Panel Patient Information Sheet
• Acute Promyelocytic Leukemia: Guideline to Diagnosis and Follow-up

Fluorescence In Situ Hybridization (FISH)

Provide a reason for referral 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.

Forms:

1. Cytogenetics Hematologic FISH Panel Patient Information Sheet (Supply T603) in Special Instructions

2. If not ordering electronically, submit a Cytogenetics Hematologic Disorders Request Form (Supply T607) with the specimen.

Advise Express Mail or equivalent if not on courier service.

Submit only 1 of the following specimens:

Specimen Type: Whole blood

Container/Tube: Green top (sodium heparin)

Specimen Volume: 7-10 mL                                          

Collection Instructions:

1. Invert several times to mix blood.

2. Other anticoagulants are not recommended and are harmful to the viability of the cells.

Specimen Type: Bone marrow

Container/Tube: Green top (sodium heparin)

Specimen Volume: 1-2 mL

Collection Instructions:

1. Invert several times to mix bone marrow.

2. Other anticoagulants are not recommended and are harmful to the viability of the cells.

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 subtypes of AML have been recognized (termed AML-M0, M1, M2, M3, M4, M5, M6, and M7) based on the cell morphology and myeloid lineage involved.

In addition to morphology, several recurrent chromosomal abnormalities have been linked to specific subtypes of AML. The most common chromosome abnormalities associated with AML include t(8;21), t(15;17), inv(16), +8, t(6;9), t(8:16), t(1;22), t(9;22) and abnormalities of the MLL gene at 11q23. The most common genes juxtaposed with MLL through translocation events in AML include MLTT4- t(6;11), MLLT3- t(9;11), MLLT10- t(10;11), CREBBP- t(11;16), ELL- t(11;19p13.1), and MLLT1-t(11;19p13.3).

AML can also evolve from myelodysplasia (MDS). Thus, the common chromosome abnormalities associated with MDS can also be identified in AML, which include: inv(3), -5/5q-, -7/7q-, +8, 13q-, 17p-, 20q-, t(1;3), and t(3;21).

In combination, the multiple 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, this analysis requires dividing cells, takes 5 to 7 days to process, and some of the subtle rearrangements can be missed (eg, inv[16] and MLL anomalies). Thus, we have validated a combination of commercially available and Mayo in-house developed FISH probes to detect the common chromosome abnormalities observed in AML patients. These probes have diagnostic and prognostic relevance and can also be used to track response to therapy.

Our panel of multiple FISH probes can be utilized to study nonproliferating (interphase) cells and can identify the all the common cytogenetic abnormalities associated 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.

Detection of an abnormal clone likely indicates a diagnosis of an acute myeloid leukemia of various subtypes.

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

This test is not approved by the FDA and it is best used as an adjunct to existing clinical and pathologic information.

Each probe was independently tested and verified on unstimulated peripheral blood and bone marrow specimens. Normal cutoff were calculated based on the results 25 normal specimens. For each probe set a series of chromosomally abnormal specimens were evaluated to confirm each probe set detected the anomaly it was designed to detect.

1. Grimwade D, Hills RK, Moorman AV, et al: Refinement of cytogenetics classification in acute myeloid leukemia: determination of prognostic significance or rare recurring chromosomal abnormalities among 5879 younger adult patients treated in the United Kingdom Research Council trials. Blood 2010 Jul;116(3):354-365

2. International Agency for Research on Cancer (IARC): World Health Organization (WHO) classification of tumour of haematopoietic and lymphoid tissues. Edited by SH Swerdlow, E Campo, NL Harris, et al. IARC Press, Oxford: Oxford University Press (distributor), 2008

Identification of deletions or monosomy of chromosomes 5, 7, 13, trisomy of chromosome 8 and deletions or rearrangements of chromosome 17 and 20 are detected based on FISH enumeration strategy probes. Rearrangements involving MLL are detected using a dual-color break-apart (BAP) strategy probe. Dual-color dual-fusion (D-FISH) strategy probe sets are used to detect inv(3) MYH11/CBFB, inv(16) RPN1/MECOM, t(8;21), t(15;17), t(6;9), t(8:16), t(3:21), t(1;3), t(11;22), t(9;22) and in reflex testing when rearrangements of the MLL gene are detected. For enumeration and BAP strategy probe sets 200 interphase nuclei are scored and 500 interphase nuclei are scored when D-FISH probes are used. Two technologists analyze each probe set and all results are expressed as percent abnormal nuclei.(Unpublished Mayo method)

Samples processed Monday through Sunday. Results reported Monday through Friday, 8 a.m.-5 p.m. CST.

Acute Myeloid Leukemia (AML), FISH

88271 x 2-DNA probe, each

88275 x 2-Interphase in situ hybridization

88291-Interpretation and report

One Additional FISH Probe

88271-DNA probe, each (if appropriate)

88275-Interphase in situ hybridization (if appropriate) 

Two Additional FISH Probes

88271 x 2-DNA probe, each (if appropriate)

88275-Interphase in situ hybridization (if appropriate) 

Three Additional FISH Probes

88271 x 3-DNA probe, each (if appropriate)

88275-Interphase in situ hybridization (if appropriate) 

Four Additional FISH Probes

88271 x 4-DNA probe, each (if appropriate)

88275 x 2-Interphase in situ hybridization (if appropriate) 

Five Additional FISH Probes

88271 x 5-DNA probe, each (if appropriate)

88275 x 2-Interphase in situ hybridization (if appropriate) 

Six Additional FISH Probes

88271 x 6-DNA probe, each (if appropriate)

88275 x 3-Interphase in situ hybridization (if appropriate) 

Seven Additional FISH Probes

88271 x 7-DNA probe, each (if appropriate)

88275 x 3-Interphase in situ hybridization (if appropriate) 

Eight Additional FISH Probes

88271 x 8-DNA probe, each (if appropriate)

88275 x 4-Interphase in situ hybridization (if appropriate) 

Nine Additional FISH Probes

88271 x 9-DNA probe, each (if appropriate)

88275 x 4-Interphase in situ hybridization (if appropriate) 

Ten Additional FISH Probes

88271 x 10-DNA probe, each (if appropriate)

88275 x 5-Interphase in situ hybridization (if appropriate) 

Eleven Additional FISH Probes

88271 x 11-DNA probe, each (if appropriate)

88275 x 5-Interphase in situ hybridization (if appropriate) 

Twelve Additional FISH Probes

88271 x 12-DNA probe, each (if appropriate)

88275 x 6-Interphase in situ hybridization (if appropriate) 

Thirteen Additional FISH Probes

88271 x 13-DNA probe, each (if appropriate)

88275 x 6-Interphase in situ hybridization (if appropriate) 

Fourteen Additional FISH Probes

88271 x 14-DNA probe, each (if appropriate)

88275 x 7-Interphase in situ hybridization (if appropriate) 

Fifteen Additional FISH Probes

88271 x 15-DNA probe, each (if appropriate)

88275 x 7-Interphase in situ hybridization (if appropriate) 

Sixteen Additional FISH Probes

88271 x 16-DNA probe, each (if appropriate)

88275 x 8-Interphase in situ hybridization (if appropriate) 

Seventeen Additional FISH Probes

88271 x 17-DNA probe, each (if appropriate)

88275 x 8-Interphase in situ hybridization (if appropriate) 

Eighteen Additional FISH Probes

88271 x 18-DNA probe, each (if appropriate)

88275 x 9-Interphase in situ hybridization (if appropriate) 

Nineteen Additional FISH Probes

88271 x 19-DNA probe, each (if appropriate)

88275 x 9-Interphase in situ hybridization (if appropriate) 

Twenty Additional FISH Probes

88271 x 20-DNA probe, each (if appropriate)

88275 x 10-Interphase in situ hybridization (if appropriate)

Twenty One Additional FISH Probes

88271 x 21-DNA probe, each (if appropriate)

88275 x 10-Interphase in situ hybridization (if appropriate)

Twenty Two Additional FISH Probes

88271 x 22-DNA probe, each (if appropriate)

88275 x 11-Interphase in situ hybridization (if appropriate)

Twenty Three Additional FISH Probes

88271 x 23-DNA probe, each (if appropriate)

88275 x 11-Interphase in situ hybridization (if appropriate)

Twenty Four Additional FISH Probes

88271 x 24-DNA probe, each (if appropriate)

88275 x 12-Interphase in situ hybridization (if appropriate)

Twenty Five Additional FISH Probes

88271 x 25-DNA probe, each (if appropriate)

88275 x 12-Interphase in situ hybridization (if appropriate)

Twenty Six Additional FISH Probes

88271 x 26-DNA probe, each (if appropriate)

88275 x 13-Interphase in situ hybridization (if appropriate)

Twenty Seven Additional FISH Probes

88271 x 27-DNA probe, each (if appropriate)

88275 x 13-Interphase in situ hybridization (if appropriate)

Twenty Eight Additional FISH Probes

88271 x 28-DNA probe, each (if appropriate)

88275 x 14-Interphase in situ hybridization (if appropriate)

Twenty Nine Additional FISH Probes

88271 x 29-DNA probe, each (if appropriate)

88275 x 14-Interphase in situ hybridization (if appropriate)

Thirty Additional FISH Probes

88271 x 30-DNA probe, each (if appropriate)

88275 x 15-Interphase in situ hybridization (if appropriate)