Abstract
BackgroundThe World Health Organization (WHO) classification system defines recurrent chromosomal translocations as the sole diagnostic and prognostic criteria for acute leukemia (AL). These fusion transcripts are pivotal in the pathogenesis of AL. Clinical laboratories universally employ conventional karyotype/FISH to detect these chromosomal translocations, which is complex, labour intensive and lacks multiplexing capacity. Hence, it is imperative to explore and evaluate some newer automated, cost-efficient multiplexed technologies to accommodate the expanding genetic landscape in AL.Methods“nCounter® Leukemia fusion gene expression assay” by NanoString was employed to detect various fusion transcripts in a large set samples (n = 94) utilizing RNA from formalin fixed paraffin embedded (FFPE) diagnostic bone marrow biopsy specimens. This series included AL patients with various recurrent translocations (n = 49), normal karyotype (n = 19), or complex karyotype (n = 21), as well as normal bone marrow samples (n = 5). Fusion gene expression data were compared with results obtained by conventional karyotype and FISH technology to determine sensitivity/specificity, as well as positive /negative predictive values.ResultsJunction probes for PML/RARA; RUNX1-RUNX1T1; BCR/ABL1 showed 100 % sensitivity/specificity. A high degree of correlation was noted for MLL/AF4 (85 sensitivity/100 specificity) and TCF3-PBX1 (75 % sensitivity/100 % specificity) probes. CBFB-MYH11 fusion probes showed moderate sensitivity (57 %) but high specificity (100 %). ETV6/RUNX1 displayed discordance between fusion transcript assay and FISH results as well as rare non-specific binding in AL samples with normal or complex cytogenetics.ConclusionsOur study presents preliminary data with high correlation between fusion transcript detection by a throughput automated multiplexed platform, compared to conventional karyotype/FISH technique for detection of chromosomal translocations in AL patients. Our preliminary observations, mandates further vast validation studies to explore automated molecular platforms in diagnostic pathology.Electronic supplementary materialThe online version of this article (doi:10.1186/s13000-016-0541-z) contains supplementary material, which is available to authorized users.
Highlights
The World Health Organization (WHO) classification system defines recurrent chromosomal translocations as the sole diagnostic and prognostic criteria for acute leukemia (AL)
In acute myeloid leukemia (AML) and acute lymphoblastic leukemia (ALL), chromosomal abnormalities play a critical role in pathogenesis and prognosis [1]
We employed adequate representation of samples with recurrent chromosomal translocation in AML (n = 23) and ALL (n = 26) while AML with normal cytogenetics (n = 19), or complex cytogenetics (n = 21) and normal bone marrow (n = 5) samples were used as negative controls
Summary
The World Health Organization (WHO) classification system defines recurrent chromosomal translocations as the sole diagnostic and prognostic criteria for acute leukemia (AL) These fusion transcripts are pivotal in the pathogenesis of AL. Specific recurrent chromosomal translocations and their fusion transcripts define the current WHO classification system for acute leukemia (AL) [3] These genetic aberrations in combination with morphology, immunophenotype and clinical features are used to diagnose distinct types of AL and thereby dictate therapeutic approaches [4]. Clinical application of RT-PCR is narrow due to variation in breakpoints resulting in several splice regions [7] Given these limitations and the expanding new discoveries of additional recurrent chromosomal translocations, there is a need to explore newer automated technologies to “accommodate” new fusion transcript discoveries and make this significant clinical diagnostic testing cost-effective and time efficient
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