Quantitative Reverse Transcription Polymerase Chain Reaction
Quantitative Reverse Transcription Polymerase Chain Reaction
Reverse Transcription Polymerase Chain Reaction
Reverse Transcription Polymerase Chain Reaction
Use for diagnosis, prognosis, and management. Not intended for MRD monitoring.
Capillary Electrophoresis
Polymerase Chain Reaction/Sequencing
Use for prognostication in cytogenetically normal AML (CN-AML).
Polymerase Chain Reaction/Sequencing
Use for prognostication in core-binding factor-related (CBF) AML.
Capillary Electrophoresis
Related Next Generation Sequencing Tests
Massively Parallel Sequencing
Massively Parallel Sequencing
Acute myeloid leukemias (AMLs) are a heterogeneous group of disorders characterized by the clonal expansion of myeloid precursors in the peripheral blood, bone marrow, and/or other tissues, which results in impaired hematopoiesis and bone marrow failure. AML is the most common acute leukemia in adults (~80% of leukemia cases) and accounts for the largest number of annual deaths from leukemia in the United States. Gene alterations, along with translocations and inversions, carry prognostic importance in AML. In addition to large chromosomal rearrangements, molecular changes have also been implicated in the development of AML. A comprehensive evaluation of several molecular markers, including FLT3, NPM1, CEBPA, KIT, IDH1, and IDH2, is important for risk assessment and prognostication in certain patients with AML, and may guide treatment decisions.
For more information on next generation sequencing testing for AML, see Myeloid Malignancies Mutation Panel by Next Generation Sequencing.
Testing Strategy
At diagnosis, the minimum AML workup includes a bone marrow aspirate for morphology, flow cytometric immunophenotyping, cytogenetics (eg, karyotyping and fluorescence in situ hybridization [FISH]), and appropriate molecular genetic testing.
Disease Overview
Incidence
>20,0000 cases/year in the U.S.
Age of Onset
Symptoms
- Symptoms resulting from thrombocytopenia, neutropenia, and anemia due to the accumulation of blasts in the marrow
- Morphologic hallmark: excessive accumulation of blasts (typically >20%) and other defined immature cells which affect one or more myeloid lineage
Test Interpretation
For more detailed information on the prognostic significance of molecular markers in AML, see the ARUP Consult Acute Myeloid Leukemia topic.
Sensitivity/Specificity
Gene | Methodology | Analytical Sensitivity | Analytical Specificity (%) |
---|---|---|---|
CEBPA |
PCR/sequencing |
40% mutated cells |
100 |
FLT3 ITD and TKD |
PCR/CE |
Signal ratio of 0.05 for ITD and 0.05 for TKD D835 |
100 |
IDH1 and IDH2 |
PCR/sequencing |
40% mutated cells |
100 |
KIT |
PCR/fragment analysis/sequencing |
30% mutated cells for exon 17 5% mutated cells for exon 8 |
100 |
NPM1 |
Quantitative reverse transcription PCR |
1:100,000 |
100 |
CBFB-MYH11a |
Quantitative reverse transcription PCR |
1:10,000 |
100 |
PML-RARAa |
Quantitative reverse transcription PCR |
1:10,000 |
85 |
RUNX1-RUNX1T1a |
Quantitative reverse transcription PCR |
1:100,000 |
100 |
aThese fusions can initially be screened by FISH but are also useful in monitoring for MRD. CE, capillary electrophoresis; PCR, polymerase chain reaction |
Limitations
- Variants outside the targeted regions or below the limit of detection will not be identified
- Results must always be interpreted within the patient's clinical context and in conjunction with other relevant data and should not be used alone for a diagnosis of malignancy
References
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28225303
Arber DA, Borowitz MJ, Cessna M, et al. Initial diagnostic workup of acute leukemia: guideline from the College of American Pathologists and the American Society of Hematology. Arch Pathol Lab Med. 2017;141(10):1342-1393.
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NCCN - Acute Myeloid Leukemia
National Comprehensive Cancer Network. NCCN Clinical Practice Guidelines in Oncology: Acute myeloid leukemia. Version 3.2020. [Last update: Dec 2019; Accessed: Sep 2020]
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28066929
Weinberg OK, Sohani AR, Bhargava P, et al. Diagnostic work-up of acute myeloid leukemia. Am J Hematol. 2017;92(3):317-321.
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27367478
De Kouchkovsky I, Abdul-Hay M. Acute myeloid leukemia: a comprehensive review and 2016 update. Blood Cancer J. 2016;6(7):e441.
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WHO Classification of Tumours of Haematopoietic and Lymphoid Tissues, Rev 4th 2017
Swerdlow S, Campo E, Jaffe E, et al. WHO Classification of Tumours of Haematopoietic and Lymphoid Tissues. 4th ed. International Agency for Research on Cancer; 2017.
Detect and quantitate gene alterations/translocations/
inversions. Use for minimal residual disease (MRD) and relapse risk monitoring.