Includes all controls
Glioblastoma Molecular Markers
Glioblastoma multiforme (GBM) represents the deadliest form of brain tumor in humans, with current treatments usually providing only minor benefit in overall survival. Recent advances in understanding molecular subtypes have led to the identification of a number of biomarkers for a more accurate classification of GBM. These molecular markers show potential for better diagnosis and prognosis of GBM patients. For example, Isocitrate Dehydrogenase 1/2 (IDH1/2) mutations have been identified mostly in secondary GBM and recently shown to carry strong favorable prognostic significance. Furthermore, evidence shows that IDH mutations are tightly associated with MGMT promoter methylation. MGMT promoter methylation leads to increased sensitivity to alkylating agents such as temozolomide and has prognostic significance similar to IDH mutations. EGFRvIII, the most common EGFR mutation in GBM, is found in approximately 60-70% of EGFR-amplified primary GBM tumors and is a potential target for experimental therapies.
EntroGen offers three (3) GBM molecular diagnostic kits for testing of IDH mutations, MGMT promoter methylation, and EGFRvIII amplification, respectively. All three assays are polymerase chain reaction (PCR)-based and use allele-specific primers in a multiplex reaction to identify their target. The MGMT Methylation Detection Kit works by amplifying the epigenetically silenced MGMT promoter regions after bisulfite treatment of DNA to determine the methylation status of the MGMT promoter. The IDH1/2 assay works by amplifying mutant-specific sequences in DNA samples. The EGFRvIII Detection Kit provides reagents that detect this mutation in total RNA isolated from tumor biopsies in a single step process (RNA to results in one step). Each kit is supplied with primer/probe sets for multiplexed detection of both target and internal control genes.
Testing Procedure and Analysis
The testing procedure involves the following simple steps:
1. Isolation of DNA or RNA from tumor biopsies, paraffin-embedded sections (FFPE), or fresh frozen tumors. (For the MGMT promoter methylation assay, the extracted DNA first undergoes bisulfite treatment.)
2. Amplification using the provided reagents in the kit.
3. Data analysis and interpretation using real-time PCR software.
Equipment & Materials
EntroGen’s GBM assays require a real-time PCR instrument capable of detecting FAM and VIC fluorescent probes.
The tests include reagents required for PCR amplification/detection, as well as validated reaction controls. Columns and reagents for DNA isolation and DNA bisulfite treatment (MGMT assay only) are not included.