Medicina Molecular e Genética

Detection of IDH1 and IDH2 Mutations in Patients with Acute Myeloid Leukemia Using Novel, Highly Sensitive Real-Time PCR Assays with Rapid Turnaround Time

Abstract

Dash DP, Wise L, Knier A, Boretsky M, Simons J, Berchanskiy D, Indig MA and Joseph AM

Isocitrate dehydrogenase 1 and 2 (IDH1/IDH2) mutations are frequent in acute myeloid leukemia (AML). Here we describe the qualitative polymerase chain reaction (PCR)-based Abbott Realtime IDH1 and IDH2 assays, which detect single nucleotide variants coding for five IDH1 and nine IDH2 mutations. We evaluated the sensitivity and specificity of the Abbott Real-Time IDH1/IDH2 assays and conducted a workflow analysis that compared them with PCR-based Sanger and next-generation sequencing (NGS) assays in blood or bone marrow specimens from 100 AML patients. Sanger sequencing and NGS detected IDH mutations with variant allele sensitivity limits of 20% and 10%, respectively. The Abbott Real-Time IDH1/IDH2 assays demonstrated 100% sensitivity and 95% specificity vs. Sanger sequencing and detected mutations at the 1% level. Low-level IDH2 mutations in five samples were detected by the Abbott Real-Time IDH2 assay but not by Sanger sequencing. Turnaround time (TAT) based on a workflow analysis showed Abbott Real-Time IDH1/IDH2 assay results were available in three business days vs. eight days for Sanger sequencing and 15 days for NGS. Higher sensitivity and rapid TAT for detecting IDH mutations may improve identification of patients with lower mutant-IDH burden and allow for quicker administration of the FDA-approved IDH inhibitors, Ivosidenib and Enasidenib.