RAPID IDH1 GENE MUTATION ANALYSIS FOR INTRAOPERATIVE PATHOLOGICAL DIAGNOSIS

摘要

BACKGROUND: (blind field). METHODS: DNA extracted in 15 min from glioma tissue collected during surgery was used to test for R132 point mutations of the IDH1 gene by using real-time PCR/high resolution melting analysis method (LightCycler 480 system). Normally, detecting IDH1 mutations requires about 80 min from the start of the PCR cycle. For rapid diagnosis, however, DNA extension and annealing times in the PCR cycle were reduced by half. Our results were compared with those obtained using the regular method. RESULTS: Regular analysis and rapid diagnosis analysis were used to detect IDH1 mutations in 6 glioma cases (diffuse astrocytoma, 2 cases; oligodendroglioma, 2 cases; anaplastic astrocytoma, 1 case; glioblastoma, 1 case). Both methods produced the same results in all cases. CONCLUSIONS: Direct DNA sequencing and immunostaining can be used to identify IDH1 mutations. However, because analyses by using the former method require several hours, it cannot be used in rapid diagnosis. The latter method could not identify all R132 mutations, because the current commercially available antibodies can only detect R132H mutant proteins. This method can detect IDH1 mutation in even 1% of tumor purity, and determine all IDH1 R132 mutation-positive gliomas during surgery in 50–60 min after the tissue is collected. Further, this method could aid in intraoperative pathological diagnoses to differentiate IDH1 mutation-positive low-malignancy gliomas from gliosis and other non-neoplastic tissues. SECONDARY CATEGORY: Clinical Neuro-Oncology.