DEVELOPMENT OF A REAL-TIME PCR MGMT PROMOTER METHYLATION ASSAY

Abstract

O6-methylguanine methyltransferase (MGMT) promoter methylation is associated with prolonged survival rates in patients with glioblastoma multiforme (GBM) undergoing standard therapy with alkylating agent Temozolomide (TMZ). Determining the MGMT methylation status of a patient diagnosed with GBM is crucial to triage appropriate clinical therapies. In this study, a real-time PCR assay was developed to detect the percent methylation reference (PMR) using designed primers and probes specific to methylated MGMT. The assay was developed using methylated and unmethylated commercial controls and cell lines to test specificity, sensitivity and limit of detection to further assess the assays robustness, efficacy and efficiency. This study shows this real-time PCR assay is effective and robust in determining the MGMT methylation status of a patient sample to aid physicians in their decision making for clinical therapies.