Impact of PCR-based molecular analysis in daily diagnosis for the patient with gliomas

Abstract

The WHO2016 CNS update requires a combined histological and molecular assessment. To assess the major aberrations such as co-deletion of complete chromosome arms 1p and 19q (Co-del), isocitrate dehydrogenase and histone H3 mutations, direct sequencing, multiplex ligation-dependent probe amplification and/or FISH are methods considered to be "golden standard" in the community. However, these methods are expensive and complicated. The aim of this study is verification of the sensitivity of the simple PCR-based techniques for assessment of molecular information in daily diagnosis. We analyzed a total number of 80 patients with gliomas. FISH and PCR-based microsatellite analysis were compared for Co-del assessment. Direct sequencing and qPCR using hig-resolution melting (HRM) were compared for IDH and histone H3 mutations. The sensitivity and specificity of FISH were 0.71 and 0.79, respectively. FISH using a commercially available Vysis probe had a risk of high false-positive rate (0.25). For assessment of IDH1 mutations, the sensitivity and specificity of HRM were 1.0 and 0.96, respectively. For assessment of IDH2 and H3 mutations by HRM, both sensitivity and specificity were 1.0. We consider PCR-based molecular analysis to be a simple and accurate technique in daily diagnosis that is readily available for a small scientific facility.

Diagnosis of Pseudoprogression Following Lomustine–Temozolomide Chemoradiation in Newly Diagnosed Glioblastoma Patients Using FET-PET

“…When immunostaining for IDH1-R132H remained negative, the mutational hotspots at codon 132 of IDH1 and codon 172 of IDH2 were directly sequenced, as reported previously (42,43). The 1p/19q codeletion status was determined by PCR-based microsatellite analysis as reported (44). The O 6 -methylguanine-DNAmethyltransferase (MGMT) promoter methylation status was assessed by methylation-specific PCR as described elsewhere (43).…”

Prediction of survival in patients with IDH-wildtype astrocytic gliomas using dynamic O-(2-[18F]-fluoroethyl)-l-tyrosine PET

“…When immunostaining for IDH1-R132H remained negative, the mutational hot-spots at codon 132 of IDH1 and codon 172 of IDH2 were directly sequenced as reported [10,38]. The 1p/19q co-deletion status was determined by PCR-based microsatellite analysis as reported [39]. The O 6 -methylguanine-DNA-methyltransferase (MGMT) promoter methylation status was assessed by methylation-specific PCR as described elsewhere [38].…”

Importance of Age and Noncontrast-Enhancing Tumor as Biomarkers for Isocitrate Dehydrogenase–Mutant Glioblastoma: A Multicenter Study

“…We retrospectively collected clinical and MRI data of 347 patients with glioblastoma in the 2016 WHO classification who preoperatively underwent 1.5- or 3-T MRI studies from 22 facilities from April 2013 to March 2017. The inclusion criteria were as follows: ( a ) patients older than 20 years; ( b ) newly diagnosed glioblastoma on histopathology; ( c ) diagnosis of IDH-wildtype or IDH-mutant glioblastoma according to the 2016 WHO classification by immunohistochemistry for IDH1 R132H mutation, high-resolution melting analysis 19 and/or IDH1/2 sequencing; ( d ) patients with clinical information, including age, sex, height, weight, symptoms, and MRI data; and ( e ) patients who preoperatively underwent conventional 1.5- or 3-T MRI studies, including diffusion-weighted imaging (DWI). The analysis details of IDH mutation status are given in the online Appendix (http://links.lww.com/RCT/A164).…”