Mutations in IDH1/2 genes are a marker of good prognosis for glioma patients, associated with low grade gliomas and secondary glioblastomas. Immunohistochemistry and Sanger sequencing are current standards for IDH1/2 genotyping while many other methods exist. The aim of this study was to validate Competitive amplification of differentially melting amplicons (CADMA) PCR for IDH genotyping by comparison with SNaPshot assay and two immunohistochemical methods. In our study, 87 glioma patients (46 from Olomouc and 41 from Ostrava) were analyzed. IDH1/2 mutations in native bioptical samples were analyzed at DNA level by CADMA and SNaPshot while IDH1 mutations in FFPE samples were analyzed at protein level by two IHC methods. CADMA PCR sensitivity for IDH1 was 96.4% and specificity 100% for 86 concluded samples. SNaPshot assay sensitivity was 92.9% and specificity of 100% for 85 concluded samples. IHC in the laboratory no. 2 reached sensitivity 85.7% and specificity 100% for 86 concluded samples. IHC in the laboratory no. 4 reached sensitivity of 96.4% and specificity of 79.7% in 74 concluded samples. Only one IDH2 mutation was found by SNaPshot while CADMA yielded false negative result. In conclusion, CADMA is a valid method for IDH1 p.(R132H) testing with higher sensitivity than SNaPshot assay. Also, molecular genetic methods of IDH1 testing from native samples were more robust than IHC from FFPE.
telomerase activity and expression level of hTERT. In the present study, we find out the effect of sodium selenite and selenium l methionine and cadmium chloride on telomerase activity in adenocarcinoma cancer cell line (AGS). Methods: The adenoma carcinoma gastric cancer cells (AGS) were cultured in RPMI1640 supplemented with 10 % fetal bovine serum (FBS) and 1% pen/strep. Concentrations of cadmium chloride, selenium l methionine and sodium selenite were added the adenocarcinoma gastric cancer cells (AGS). Finally, the telomerase activity was measured by telomeric repeat amplification protocol (TRAP) and qRT-TRAP assays. Results: Average telomerase activity (as threshold cycle (CT)) has been detected for three concentrations (5lM and 10lM and 20 lM) of the noted compounds. The results were 24.
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