Molecular diagnosis of glioma by PCR amplification technology

Ref: Yokogami K, et alBrain Brain Tumor Pathol2018 Jun 21doi: 10.1007/s10014-018-0322-3The 2016 edition of the
WHO Central Nervous System (CNS) Tumor Classification states that IT is necessary to diagnose CNS tumors in combination with histology and genotypeAt present, direct sequencing, multi-connecting probe amplification technology (MLPA) or fluorescence in situ hybridization (FISH) are important means of detecting chromosomal 1p/19q colossus (Co-del), isocytic citric acid dehydrogenase (IDH) and histone H3 mutationsHowever, these methods are complex and expensiveKiyotaka Yokogami of the University of Japan's Miyazaki University School of Medicine, Japan, and others used a simple polymerase chain reaction amplification (PCR amplification) technique for molecular diagnosis of gliomasThe results were published online in June 2018 in Brain Pathologythe authors compared the results of 1p/19q Co-del with FISH and PCR-based microsatellite analysis (PCR-based microsatellite analysis) in 80 glioma specimens, and quantitative polymerase chain reaction (qPCR) and direct sequencing compared IDH and histone H3 mutations using high-resolution melting curve analysis (HRM)found that 7 out of 61 patients were diagnosed as Co-del through microsatellite analysis, and that the effect of using FISH to evaluate Co-del was significantly differentTHE SENSITIVITY AND SPECIFICITY OF FISH WERE 0.71 AND 0.79, AND THE FALSE POSITIVE AND NEGATIVE RATES WERE 0.21 AND 0.29, RESPECTIVELYin the trial of idh R132H mutation seyness with immunohiscita (IHC) staining, 2 of the 31 patients were falsely positive, with sensitivity and specificity of 1.0 and 0.92, respectivelyin HRM analysis, in order to avoid false negatives, the following assumptions are made: when both amplification results are mutant, it is considered mutant, and when both amplification results are wild, it is considered wildIn the first analysis, the results of 5 cases (8%) glioma samples were not determined, and in the second analysis, the results of these 5 samples were stableOf the 60 tumor samples, 2 (3.3%) were falsely positiveOverall, hrM detected the sensitivity and specificity of THE IDH1 mutation, respectively, 1.0 and 0.96 When HRM detected the mutation of IDH2, in 31 tumor samples, 1 case had a difference in the recognition of HRM, sensitivity and specificity were 1.0 Three H3F3A and one HIST1H3B mutation can also be detected by HRM, with both specificity and sensitivity of 1.0 Low-quality DNA was extracted from Falmarin's fixed paraffin (FFPE) samples for detection, and there were some differences in waveforms, but the results were not different and the results were satisfactory , the author points out that PCR-based amplification technology can be a satisfactory molecular diagnosis of glioma, which is simple, accurate and feasible (
Xuan compiled, Fudan University affiliated Huashan Hospital Huashan Hospital, Dr Huayu review, "Outside the Information" editor-in-chief, Fudan University affiliated Huashan Hospital Chen Rongcheng Professor Final Review) related links