What is IDH1/2?

IDH1 and IDH2 are genes encoding isocitrate dehydrogenase enzymes involved in cellular metabolism. These enzymes catalyze the oxidative decarboxylation of isocitrate to alpha-ketoglutarate (α-KG). IDH1 operates in the cytoplasm, while IDH2 functions in the mitochondria. Mutations in IDH1 (commonly R132) and IDH2 (commonly R140 and R172) are somatic and result in neomorphic enzymatic activity, leading to the accumulation of the oncometabolite D-2-hydroxyglutarate (2-HG), a competitive inhibitor of α-KG-dependent enzymes.

Role of IDH1/2 Mutation in Disease

IDH1/2 mutations are found in a range of malignancies, including:

• Acute myeloid leukemia (AML): Present in ~7–14% (IDH1) and 8–19% (IDH2) of AML cases, especially in patients with normal karyotypes.

• Low-grade gliomas and secondary glioblastomas: Over 80% prevalence of IDH mutations.

These mutations affect epigenetic regulation by disrupting histone demethylation and DNA hydroxymethylation, contributing to abnormal cell proliferation, impaired differentiation, and treatment resistance. Detecting IDH1/2 mutations is essential for diagnosis, prognosis, and emerging targeted therapies (e.g., IDH inhibitors).

About the TRUPCR® IDH1/2 Mutation Detection Kit

The TRUPCR® IDH1/2 Mutation Detection Kit is an in vitro diagnostic (IVD) assay designed to detect common somatic mutations in the IDH1 and IDH2 genes. It uses allele-specific amplification via ARMS PCR combined with fluorescent probe detection in a Real-Time PCR platform.

Target samples:

• Human blood (EDTA)

• Bone marrow (EDTA)

Detected mutations include:

• IDH1 R132

• IDH1 R100

• IDH2 R140

• IDH2 R172

Included in the kit:

• Mutation-specific reaction mix

• Reference control mix

• Internal control

• Positive and negative controls

All components are provided in a ready-to-use format, ensuring a streamlined workflow in clinical laboratories.

Principle and Workflow

The kit is based on the ARMS PCR method. The assay uses allele-specific primers that allow selective amplification of mutant DNA in a background of predominantly wild-type DNA. The amplification is monitored via fluorescent-labeled probes using FRET-based detection.

Workflow overview:

1. DNA extraction from blood or bone marrow

2. Setup of the Real-Time PCR reaction with multiplex tubes

3. Thermal cycling and fluorescence data collection

4. Mutation analysis based on cycle threshold (Ct) values

Key benefits of the method:

• High specificity for mutations

• Internal control for reaction validity

• No post-PCR handling

• Suitable for high-throughput testing

Clinical Application

The TRUPCR® IDH1/2 Mutation Detection Kit provides essential data for the molecular classification of AML and other IDH-driven malignancies. Its application includes:

• Risk stratification and prognostic assessment in AML

• Patient eligibility for targeted therapy (e.g., Enasidenib, Ivosidenib)

• Monitoring mutation burden during treatment

• Support for research and clinical studies in hematologic malignancies

Its CE-IVD certification ensures it meets EU regulatory requirements for routine clinical diagnostics.