FISH – PANELS*

Fluorescence In Situ Hybridization – AML/MDS Panel

Test Overview

CPT Code(s)

88237-52, 88271×3, 88275, 88291

Methodology

Culture/Hybridization/Microscopy/Interpretation

Specimen Requirements

Bone Marrow and/or Leukemia Blood

Container: Sodium Heparin (green top tube)

Optimal Quantity: 2-3 ml

Minimum Quantity: 1-2 ml

Storage: Room Temperature

Stability at Room Temperature: 8 hours, then refrigerate

Transportation: Avoid freezing or heating over 35oC

Turnaround Time

Final report in 5 days for 90% of cases

Test Details

Clinical Significance

Fluorescence in situ hybridization (FISH) is a sensitive method to detect smaller genomic changes associated with various hematological malignancies and solid tumors.  There are several advantages with FISH technology over routine chromosome analysis and such advantages include the ability of FISH technology to detect genomic abnormalities in non-viable and non-dividing tissues, rapid turnaround time, and increased resolution.  However, FISH technology is complementary to routine chromosome analysis and cannot substitute routine chromosome analysis for diagnosis of cancer.

Clinical Background

Acute Myelogenous Leukemia/Myelodysplastic syndrome is a group of heterogeneous hematologic conditions with characteristic chromosome abnormalities leading to specific gene rearrangements.  About half of the patients with AML/MDS may show chromosome abnormalities which can be classified as primary and secondary or treatment related abnormalities.  The chromosome abnormalities are known to correlate with prognosis and response to treatment.  Although majority of such abnormalities can be detected on routine chromosome analysis, interphase FISH analysis due to its high sensitivity can detect low-level clones thus identifying prognostically and therapeutically important genetic abnormalities.

Methods

The samples are usually cultured for 24-hours before hybridizing with the probes.  After hybridization and post-hybridization washes, the slides are analyzed under a fluorescence microscope equipped with epifluorescence and appropriate filter sets.  Analysis and scoring is carried out by two certified technologists in a blinded fashion.  Results are reviewed by both the laboratory manager and the director. The AML/MDS FISH panel includes the following probes.

5q deletion / monosomy 5 TAS2R1 / EGR1 Cytocell (cat # LPH024)
7q deletion / monosomy 7 RELN / TES Cytocell (cat # LPH025)
t(8;21) ETO / AML1 (RUNX1/RUNX1T1) Cytocell (cat # LPH026)
11q rearrangement MLL Cytocell (cat # LPH013)
t(15;17) PML / RARA Cytocell (cat # LPH023)
inv(16) MYH11 / CBFB Cytocell (cat # LPH022)
17p deletion p53 Cytocell (cat # LPH017)
20q deletion / monosomy 20 MYBL2 / PTPRT Cytocell (cat # LPH020)

Interpretation of Results

The morphologic interpretation and correlation of results on all cases is performed by a board-certified doctoral level scientist (laboratory director). The final report has a narrative description of results.  If abnormalities are present, they are explained in a paragraph which helps to clarify and correlate chromosomal findings with phenotype. References are included in the report to help the referring physician with interpretation, which include books or journals that contain appropriate information. Recommendations are made as to any additional testing, if necessary.

 

Fluorescence In Situ Hybridization – ALL Panel

Test Overview

CPT Code(s)

88237-52, 88271×3, 88275, 88291

Methodology

Culture/Hybridization/Microscopy/Interpretation

Specimen Requirements

Bone Marrow and/or Leukemia Blood

Container: Sodium Heparin (green top tube)

Optimal Quantity: 2-3 ml

Minimum Quantity: 1-2 ml

Storage: Room Temperature

Stability at Room Temperature: 8 hours, then refrigerate

Transportation: Avoid freezing or heating over 35oC

Test Details

Clinical Significance

Fluorescence in situ hybridization (FISH) is a sensitive method to detect smaller genomic changes associated with various hematological malignancies and solid tumors.  There are several advantages with FISH technology over routine chromosome analysis and such advantages include the ability of FISH technology to detect genomic abnormalities in non-viable and non-dividing tissues, rapid turnaround time, and increased resolution.  However, FISH technology is complementary to routine chromosome analysis and cannot substitute routine chromosome analysis for diagnosis of cancer.

Clinical Background

Acute lymphoblastic/lymphocytic leukemia (ALL) is the most common hematological malignancy seen in children.  A large number of specific chromosome abnormalities and/or gene rearrangements are seen in patients with ALL with the incidence of such abnormalities ranging from 65-85% for adults and 60-70% for children.  Detection of these abnormalities are vital in stratification of patients into different treatment groups and it is now mandatory to carry out chromosome and FISH studies, especially in children, for enrollment into Children’s Oncology Group (COG) treatment protocols. Although majority of such abnormalities can be detected on routine chromosome analysis, interphase FISH analysis due to its high sensitivity can detect low-level clones thus identifying prognostically and therapeutically important genetic abnormalities.

Methods

The samples are usually cultured for 24-hours before hybridizing with the probes.  After hybridization and post-hybridization washes, the slides are analyzed under a fluorescence microscope equipped with epifluorescence and appropriate filter sets.  Analysis and scoring is carried out by two certified technologists in a blinded fashion.  Results are reviewed by both the laboratory manager and the director. The ALL FISH panel includes the following probes.

8q rearrangement cMYC Cytocell (cat # LPH010)
9p deletion p16 / D9Z3 Cytocell (cat # LPH009)
hyperdiploidy – chr. 4 FIP1L1 / CHIC2 / PDGFRA Cytocell (cat # LPH032)
hyperdiploidy – chrs. 10 & 17 D10Z1 / D17Z1 Cytocell (cat # LPE010G/LPE017R)
t(12;21) ETV6 / AML1 Cytocell (cat # LPH012)
11q rearrangement MLL Cytocell (cat # LPH013)
t(9;22) ABL1 / BCR Cytocell (cat # LPH038)
14q rearrangement IGH Cytocell (cat # LPH014)

Interpretation of Results

The morphologic interpretation and correlation of results on all cases is performed by a board-certified doctoral level scientist (laboratory director). The final report has a narrative description of results.  If abnormalities are present, they are explained in a paragraph which helps to clarify and correlate chromosomal findings with phenotype. References are included in the report to help the referring physician with interpretation, which include books or journals that contain appropriate information. Recommendations are made as to any additional testing, if necessary.

 

Fluorescence In Situ Hybridization – Multiple Myeloma Panel

Test Overview

CPT Code(s)

88237-52, 88271×3, 88275, 88291

Methodology

Plasma cell isolation/Culture/Hybridization/Microscopy/Interpretation

Specimen Requirements

Bone Marrow and/or Leukemia Blood

Container: Sodium Heparin (green top tube)

Optimal Quantity: 2-3 ml

Minimum Quantity: 1-2 ml

Storage: Room Temperature

Stability at Room Temperature: 8 hours, then refrigerate

Transportation: Avoid freezing or heating over 35oC

Test Details

Clinical Significance

Fluorescence in situ hybridization (FISH) is a sensitive method to detect smaller genomic changes associated with various hematological malignancies and solid tumors.  There are several advantages with FISH technology over routine chromosome analysis and such advantages include the ability of FISH technology to detect genomic abnormalities in non-viable and non-dividing tissues, rapid turnaround time, and increased resolution.  However, FISH technology is complementary to routine chromosome analysis and cannot substitute routine chromosome analysis for diagnosis of cancer.

Clinical Background

Plasma cell disorders are genetically diverse group of diseases arising from malignant proliferation of monoclonal population of plasma cells.  This group consists of monoclonal gammopathy of undetermined significance (MGUS), plasma cytoma, smoldering myeloma, and plasma cell myeloma or multiple myeloma.  Genomic abnormalities are important prognostic factors in myeloma.  Routine chromosome anlaysis is often limiting and is not efficient in detecting these genomic abnormalities since there is limited infiltration of plasma cells in the bone marrow and also due to the slow proliferation of plasma cells.  Studies have shown enrichment for plasma cells is most efficient in detecting these prognostically significant genomic abnormalities.  Our lab has unique expertise in isolating plasma cells and using the negative fraction for routine chromosome analysis to detect non-plasma cell abnormalities.  In fact, we are the first laboratory to design a protocol utilizing the negative fraction for chromosome analysis with considerable success.

Methods

The plasma cells are isolated using a CD134 magnetic antibody separation method. The isolated plasma cells are harvested and hybridized with the probes.  After hybridization and post-hybridization washes, the slides are analyzed under a fluorescence microscope equipped with epifluorescence and appropriate filter sets.  Analysis and scoring is carried out by two certified technologists in a blinded fashion.  Results are reviewed by both the laboratory manager and the director. The multiple myeloma panel includes the following probes.

11q deletion D11Z1 / ATM Cytocell (cat # LPH011)
17p deletion p53 Cytocell (cat # LPH017)
13q deletion RB1 / CTB-163C9 Cytocell (cat # LPS011)
t(11;14) CCND1 / IGH Cytocell (cat # LPH021)
t(4;14) FGFR3 / IGH Cytocell (cat # LPH030)
t(14;16) IGH / MAF Cytocell (cat # LPH029)
hyperdiploidy ATM / MAF / FGFR3

Interpretation of Results

The morphologic interpretation and correlation of results on all cases is performed by a board-certified doctoral level scientist (laboratory director). The final report has a narrative description of results.  If abnormalities are present, they are explained in a paragraph which helps to clarify and correlate chromosomal findings with phenotype. References are included in the report to help the referring physician with interpretation, which include books or journals that contain appropriate information. Recommendations are made as to any additional testing, if necessary.

 

Fluorescence In Situ Hybridization – CLL Panel

Test Overview

CPT Code(s)

88237-52, 88271×3, 88275, 88291

Methodology

Culture/Hybridization/Microscopy/Interpretation

Specimen Requirements

Bone Marrow and/or Leukemia Blood

Container: Sodium Heparin (green top tube)

Optimal Quantity: 2-3 ml

Minimum Quantity: 1-2 ml

Storage: Room Temperature

Stability at Room Temperature: 8 hours, then refrigerate

Transportation: Avoid freezing or heating over 35oC

Test Details

Clinical Significance

Fluorescence in situ hybridization (FISH) is a sensitive method to detect smaller genomic changes associated with various hematological malignancies and solid tumors.  There are several advantages with FISH technology over routine chromosome analysis and such advantages include the ability of FISH technology to detect genomic abnormalities in non-viable and non-dividing tissues, rapid turnaround time, and increased resolution.  However, FISH technology is complementary to routine chromosome analysis and cannot substitute routine chromosome analysis for diagnosis of cancer.

Clinical Background

Chronic lymphocytic leukemia (CLL) is leukemia of small mature B-cells and mostly affect adults age 65 and above.  CLL is the most common lymphoid malignancy accounting for about 11% of all hematological malignancies and 25% of all leukemias.  Up to 50% of patients with CLL may have clonal cytogenetic abnormalities identified on routine chromosome analysis.  However, CLL clones (lymphocytes) do not divide in culture and are resistant to mitogens, thus making routine cytogenetic analysis more difficult.  FISH on the other hand is highly sensitive in detecting the genomic changes that have significant prognostic implications in CLL.

Methods

The samples are usually cultured for 24-hours before hybridizing with the probes.  After hybridization and post-hybridization washes, the slides are analyzed under a fluorescence microscope equipped with epifluorescence and appropriate filter sets.  Analysis and scoring is carried out by two certified technologists in a blinded fashion.  Results are reviewed by both the laboratory manager and the director. The CLL panel includes the following probes.

6q deletion D6Z1 / MYB Cytocell (cat # LPH016)
trisomy 12 D12Z1 Cytocell (cat # LPE012R)
11q deletion D11Z1 / ATM Cytocell (cat # LPH011)
t(14;18) IGH / BCL2 Cytocell (cat # LPH018)
14q rearrangement IGH Cytocell (cat # LPH014)
17p deletion p53 Cytocell (cat # LPH017)
t(11;14) CCND1 / IGH Cytocell (cat # LPH021)
13q deletion RB1 / CTB-163C9 Cytocell (cat # LPS011)

Interpretation of Results

The morphologic interpretation and correlation of results on all cases is performed by a board-certified doctoral level scientist (laboratory director). The final report has a narrative description of results.  If abnormalities are present, they are explained in a paragraph which helps to clarify and correlate chromosomal findings with phenotype. References are included in the report to help the referring physician with interpretation, which include books or journals that contain appropriate information. Recommendations are made as to any additional testing, if necessary.

Fluorescence In Situ Hybridization – MPD Panel

Test Overview

CPT Code(s)

88237-52, 88271×3, 88275, 88291

Methodology

Culture/Hybridization/Microscopy/Interpretation

Specimen Requirements

Bone Marrow and/or Leukemia Blood

Container: Sodium Heparin (green top tube)

Optimal Quantity: 2-3 ml

Minimum Quantity: 1-2 ml

Storage: Room Temperature

Stability at Room Temperature: 8 hours, then refrigerate

Transportation: Avoid freezing or heating over 35oC

Test Details

Clinical Significance

Fluorescence in situ hybridization (FISH) is a sensitive method to detect smaller genomic changes associated with various hematological malignancies and solid tumors.  There are several advantages with FISH technology over routine chromosome analysis and such advantages include the ability of FISH technology to detect genomic abnormalities in non-viable and non-dividing tissues, rapid turnaround time, and increased resolution.  However, FISH technology is complementary to routine chromosome analysis and cannot substitute routine chromosome analysis for diagnosis of cancer.

Clinical Background

Myeloproliferative neoplasms associated with activated tyrosine kinase and presenting with eosinophilia are a distinct group of myeloproliferative and lymphoid neoplasms.  These neoplasms, especially those with rearrangements involving the platelet derived growth factor receptor alpha (PDGFRA), platelet derived growth factor receptor beta (PDGFRB), fibroblast growth factor receptor 1 (FGFR1) and BCR/ABL1 fusion, respond well to treatment with tyrosine kinase inhibitors like imatinib and hence are called imatinib responsive neoplasms.  The most common chromosomal rearrangements seen in this group include t(4;12) resulting in PDGFRA rearrangement, t(5;12) resulting in PDGFRB rearrangement, t(9;22) resulting in BCR/ABL1 rearrangement  and chromosome 8 abnormalities resulting in FGFR1rearrangements.  Although some of these chromosome abnormalities can be detected on routine chromosome analysis, FISH is more sensitive in identifying and accurately characterizing the underlying gene rearrangements and hence is the preferred method for diagnosis.

Methods

The samples are usually cultured for 24-hours before hybridizing with the probes.  After hybridization and post-hybridization washes, the slides are analyzed under a fluorescence microscope equipped with epifluorescence and appropriate filter sets.  Analysis and scoring is carried out by two certified technologists in a blinded fashion.  Results are reviewed by both the laboratory manager and the director. The MPD panel includes the following probes.

t(9;22) ABL1 / BCR Cytocell (cat # LPH038)
4q rearrangement FIP1L1 / CHIC2 / PDGFRA Cytocell (cat # LPH032)
t(5;12) PDGFRB Cytocell (cat # LPH031)
8p11 rearrangement FGFR1 Cytocell (cat # LPS018)

Interpretation of Results

The morphologic interpretation and correlation of results on all cases is performed by a board-certified doctoral level scientist (laboratory director). The final report has a narrative description of results.  If abnormalities are present, they are explained in a paragraph which helps to clarify and correlate chromosomal findings with phenotype. References are included in the report to help the referring physician with interpretation, which include books or journals that contain appropriate information. Recommendations are made as to any additional testing, if necessary.

 

Fluorescence In Situ Hybridization – B-cell Lymphoma Panel

Test Overview

CPT Code(s)

88237-52, 88271×3, 88275, 88291

Methodology

Culture/Hybridization/Microscopy/Interpretation

Specimen Requirements

Bone Marrow and/or Leukemia Blood Lymph Node
Container: Sodium Heparin (green top tube) Container: Transport Media/Saline
Optimal Quantity: 2-3 ml Optimal Quantity: 23 cm
Minimum Quantity: 1-2 ml Minimum Quantity: 13 cm
Storage: Room Temperature Storage: Room Temperature
Stability at Room Temperature: 8 hours, then refrigerate Stability at Room Temperature: 8 hours, then refrigerate
Transportation: Avoid freezing or heating over 35oC Transportation: Avoid freezing or heating over 35o

Test Details

Clinical Significance

Fluorescence in situ hybridization (FISH) is a sensitive method to detect smaller genomic changes associated with various hematological malignancies and solid tumors.  There are several advantages with FISH technology over routine chromosome analysis and such advantages include the ability of FISH technology to detect genomic abnormalities in non-viable and non-dividing tissues, rapid turnaround time, and increased resolution.  However, FISH technology is complementary to routine chromosome analysis and cannot substitute routine chromosome analysis for diagnosis of cancer.

Clinical Background

B-cell lymphoma is a large group of heterogenous disorders and comprises a majority of non-Hodgkin lymphomas.  The most common abnormality seen in these disorders is the IGH gene translocations or rearrangements involving the 14q32 locus.  Included in this group is follicular lymphoma (FL), Burkitt Lymphoma (BL) and diffuse large B-cell lymphoma (DLBCL).  The characteristic chromosome abnormality, t(14;18) resulting in IGH/BCL2 fusion is seen in about 80% of FL, and about 20-30% of DLBCL while a majority of patients with BL harbor the characteristic t(8;14) resulting in cMYC/IGH fusion.  BCL6 gene rearrangements are also associated with B-cell lymphoma. Cyclin D1 gene rearrangements, especially the t(11;14) distinguishes mantle cell lymphoma (ML) from other lymphoproliferative disorders and hence is diagnostic for ML. Although majority of such abnormalities can be detected on routine chromosome analysis, interphase FISH analysis due to its high sensitivity can detect low-level clones thus identifying prognostically and therapeutically important genetic abnormalities.

Methods

The samples are usually cultured for 24-hours before hybridizing with the probes.  After hybridization and post-hybridization washes, the slides are analyzed under a fluorescence microscope equipped with epifluorescence and appropriate filter sets.  Analysis and scoring is carried out by two certified technologists in a blinded fashion.  Results are reviewed by both the laboratory manager and the director. The B-cell lymphoma panel includes the following probes.

t(14;18) IGH / BCL2 Cytocell (cat # LPH018)
8q rearrangement cMYC Cytocell (cat # LPH010)
3q rearrangement BCL6 Cytocell (cat # LPH035)
t(11;14) CCND1 / IGH Cytocell (cat # LPH021)

Interpretation of Results

The morphologic interpretation and correlation of results on all cases is performed by a board-certified doctoral level scientist (laboratory director). The final report has a narrative description of results.  If abnormalities are present, they are explained in a paragraph which helps to clarify and correlate chromosomal findings with phenotype. References are included in the report to help the referring physician with interpretation, which include books or journals that contain appropriate information. Recommendations are made as to any additional testing, if necessary.

 

Fluorescence In Situ Hybridization – T-cell Lymphoma Panel

Test Overview

CPT Code(s)

88237-52, 88271×3, 88275, 88291

Methodology

Culture/Hybridization/Microscopy/Interpretation

Specimen Requirements

Bone Marrow and/or Leukemia Blood Lymph Node
Container: Sodium Heparin (green top tube) Container: Transport Media/Saline
Optimal Quantity: 2-3 ml Optimal Quantity: 23 cm
Minimum Quantity: 1-2 ml Minimum Quantity: 13 cm
Storage: Room Temperature Storage: Room Temperature
Stability at Room Temperature: 8 hours, then refrigerate Stability at Room Temperature: 8 hours, then refrigerate
Transportation: Avoid freezing or heating over 35oC Transportation: Avoid freezing or heating over 35oC

 Turnaround Time

Final report in 5 days for 90% cases

Test Details

Clinical Significance

Fluorescence in situ hybridization (FISH) is a sensitive method to detect smaller genomic changes associated with various hematological malignancies and solid tumors.  There are several advantages with FISH technology over routine chromosome analysis and such advantages include the ability of FISH technology to detect genomic abnormalities in non-viable and non-dividing tissues, rapid turnaround time, and increased resolution.  However, FISH technology is complementary to routine chromosome analysis and cannot substitute routine chromosome analysis for diagnosis of cancer.

Clinical Background

The common genomic abnormalities seen in T-cell lymphoma usually involve the T-cell receptor genes (TCR) with various transcription factors.  The genes that are commonly involved are TCR-alpha (TCRA) and TCR-delta (TCRD) at 14q11, TCR-beta (TCRB) at 7q35, TCR-gamma (TCRG) at 7p15.  The ALK gene rearrangements occur in about 2% of adults and 13% of children.

Methods

The samples are usually cultured for 24-hours before hybridizing with the probes.  After hybridization and post-hybridization washes, the slides are analyzed using a fluorescence microscope equipped with epifluorescence and appropriate filter sets.  Analysis and scoring is carried out by two certified technologists in a blinded fashion.  Results are reviewed by both the laboratory manager and the director.  The T-cell lymphoma FISH panel includes the following probes.

inv(7) / iso(7q) TCRB Cytocell (cat # LPH048)
inv(14) / 14q rearrangement TCRAD Cytocell (cat # LPH047)
t(2;5) / 2p rearrangement ALK Cytocell (cat # LPS 019)

 Interpretation of Results

The morphologic interpretation and correlation of results on all cases is performed by a board-certified doctoral level scientist (laboratory director). The final report has a narrative description of results.  If abnormalities are present, they are explained in a paragraph which helps to clarify and correlate chromosomal findings with phenotype. References are included in the report to help the referring physician with interpretation, which include books or journals that contain appropriate information. Recommendations are made as to any additional testing, if necessary.

*Probes used on panels may be ordered as single tests