GEN-8 Real-time PCR based intraoperative genetic analysis for gliomas

Abstract An individual therapeutic strategy based on the genetic characterization is important in gliomas. However, it has been difficult to obtain genetic features during surgery. In this study, we present an overview of intraoperative genetic analysis using modified real-time PCR method. The tumor specimen was crushed with liquid nitrogen, then extract DNA within 60 minutes. Reagents of real-time PCR for detecting IDH, TERT, and BRAF hot spot mutations were stocked and real-time PCR was performed after mixing the extracted DNA. We used PNA and LNA to detect single nucleotide variant (SNV). The average time from tumor extraction to intraoperative tentative judgement was approximately 100 minutes. Using this system. we preliminary performed intraoperative genomic analysis in10 glioma patients. We confirmed that 8 of 10 cases (80%) of intraoperative genomic diagnosis were consistent with post-operative diagnosis by Sanger sequencing. However, we experienced 2 (20%) unmatched cases due to low allele of SNV, which indicates that more advanced system is required for clinical application.