Detection of recurrent balanced translocations in Acute Myeloid Leukemia by Multiplex RT-PCR

Authors

  • Manoj Gopal Madakshira Reader, Dept of Pathology Armed Forces Medical College Pune - 411040
  • Jyoti Kotwal Professor and Head of Department Department of Hematology Gangaram Hospital New Delhi

Keywords:

acute myeloid leukemia, multiplex, polymerase chain reaction

Abstract

Background: Diagnosis of acute myeloid leukemia (AML) has to be established by immunophenotyping and molecular testing for presence of recurrent genetic aberrations. This helps clinician to plan therapeutic protocol as AMLs are a heterogenous group of malignancies having varied response to treatment and prognosis. Multiplex Reverse Transcriptase Polymerase Chain Reaction (RT-PCR) is method capable of detecting 28 known fusion-transcripts of acute leukemia including cryptic translocations which would otherwise be missed on conventional cytogenetics.

Methods: In this case series, a total of 19 consecutive patients diagnosed as AML were included. Morphology and cytochemical character of blasts were ascertained. Immunophenotying was carried out by flowcytometry. Messenager RNA (mRNA) was extracted from peripheral blood or bone marrow aspirates and converted to Complimentary DNA (cDNA). The cDNA was then subjected to multiplex RT-PCR for presence of fusion-transcripts using primers from Hemavision kit, DNA technologies, Denmark. The amplified product was correlated with the morphological, cytochemical and immunophenotypic character of AML.

Result: Of the 19 cases of AML, 12 cases showed amplification of a fusion transcript. Most cases with fusion-transcripts showed an expected blast morphology and immunophenotype. Multiplex RT-PCR picked up presence of rare translocations such as t(16,21), t(9;22), t(6;9) and t(3;5), with unique  clinicopathological features.

Conclusion: Multiplex RT-PCR is a unique assay to identify multiple known recurrent genetic aberrations, including many splice variants. This aids in accurate subtyping of AML and helps the clinician to formulate a specific treatment plan for the patient. 

 

DOI: 10.21276/apalm.2017.1071

Author Biographies

Manoj Gopal Madakshira, Reader, Dept of Pathology Armed Forces Medical College Pune - 411040

Reader 

Department of Pathology 

Jyoti Kotwal, Professor and Head of Department Department of Hematology Gangaram Hospital New Delhi

Professor and Head of Department 

Department of Hematology 

 

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Published

19-02-2017

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1.
Madakshira MG, Kotwal J. Detection of recurrent balanced translocations in Acute Myeloid Leukemia by Multiplex RT-PCR. Ann of Pathol and Lab Med [Internet]. 2017 Feb. 19 [cited 2025 Jan. 14];4(1):A54-62. Available from: https://pacificejournals.com/journal/index.php/apalm/article/view/apalm1071

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Vol. 4 No. 1 (2017)

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Original Article

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Copyright (c) 2017 Manoj Gopal Madakshira, Jyoti Kotwal

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