Diagnostic testing for IDH1 and IDH2 variants in acute myeloid leukemia an algorithmic approach using high-resolution melting curve analysis

Keyur P Patel; Bedia A Barkoh; Zhao Chen; Deqin Ma; Neelima Reddy; L Jeffrey Medeiros; Rajyalakshmi Luthra

doi:10.1016/j.jmoldx.2011.06.004

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Diagnostic testing for IDH1 and IDH2 variants in acute myeloid leukemia an algorithmic approach using high-resolution melting curve analysis

Journal article

Open access

Peer reviewed

Diagnostic testing for IDH1 and IDH2 variants in acute myeloid leukemia an algorithmic approach using high-resolution melting curve analysis

Keyur P Patel, Bedia A Barkoh, Zhao Chen, Deqin Ma, Neelima Reddy, L Jeffrey Medeiros and Rajyalakshmi Luthra

The Journal of molecular diagnostics : JMD, Vol.13(6), pp.678-686

11/2011

DOI: 10.1016/j.jmoldx.2011.06.004

PMCID: PMC3194054

PMID: 21889610

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Abstract

Isocitrate dehydrogenase 1 (IDH1) and IDH2 mutations and polymorphism are reported in 5% to 15% of acute myeloid leukemia (AML) cases, with G105 and R132 of IDH1 and R140 and R172 of IDH2 known to be clinically significant. Current Sanger sequencing assays to detect IDH mutations are labor intensive and not cost effective for clinical testing of low-frequency mutations. Therefore, we developed clinical assays using high-resolution melting (HRM) analysis to screen for all four variants listed above, followed by Sanger sequencing confirmation. The sensitivities of the assays were 7.3% and 7.9% for the detection of IDH2 and IDH1 variants, respectively, against the background of wild-type transcripts. Comparison of HRM to Sanger sequencing on 146 AML bone marrow samples for validation showed near-perfect concordance for all positive and negative results for IDH1 (98%) and IDH2 (94%). Postvalidation clinical implementation of upfront HRM screening (N = 106), using a more conservative algorithm to avoid false-negative results, reduced the number of Sanger sequencing tests by 73% (IDH1) and 78% (IDH2). Of the variant calls made by HRM in postvalidation clinical samples, Sanger confirmed the presence of a variant in 62% (IDH1) and 44% (IDH2) of the samples. In conclusion, our HRM assays are rapid, convenient, and versatile assays for screening and confirmation of alterations in IDH1 and IDH2.

Nucleic Acid Denaturation

Humans

Genotype

Isocitrate Dehydrogenase - genetics

DNA Mutational Analysis - methods

Sequence Analysis, DNA

Algorithms

Leukemia, Myeloid, Acute - diagnosis

Base Sequence

Sensitivity and Specificity

Polymorphism, Single Nucleotide

Mutation

Leukemia, Myeloid, Acute - genetics

Details

Title: Subtitle: Diagnostic testing for IDH1 and IDH2 variants in acute myeloid leukemia an algorithmic approach using high-resolution melting curve analysis
Creators: Keyur P Patel - Department of Hematopathology, University of Texas M.D. Anderson Cancer Center, Houston, TX 77054, USA
Bedia A Barkoh
Zhao Chen
Deqin Ma
Neelima Reddy
L Jeffrey Medeiros
Rajyalakshmi Luthra
Resource Type: Journal article
Publication Details: The Journal of molecular diagnostics : JMD, Vol.13(6), pp.678-686
Publisher: United States
DOI: 10.1016/j.jmoldx.2011.06.004
PMID: 21889610
PMCID: PMC3194054
ISSN: 1525-1578
eISSN: 1943-7811
Language: English
Date published: 11/2011
Academic Unit: Pathology
Record Identifier: 9984047662602771

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