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.

CGB Laboratory Inc Ostrava Czech Republic

Department of Biomedical Sciences Faculty of Medicine University of Ostrava Ostrava Czech Republic

Department of Clinical and Molecular Pathology Faculty of Medicine and Dentistry Palacky University and University Hospital in Olomouc Olomouc Czech Republic

Department of Neurosurgery University Hospital Olomouc Olomouc Czech Republic

Department of Pathology University of Utah Salt Lake City UT USA

Institute of Molecular and Translational Medicine Faculty of Medicine and Dentistry Palacky University and University Hospital in Olomouc Hnevotinska 5 779 00 Olomouc Czech Republic

Institute of Pathology Faculty of Medicine and University Hospital University of Ostrava Syllabova 19 703 00 Ostrava Zábřeh Czech Republic

Neurosurgery Clinic University Hospital Ostrava Ostrava Czech Republic

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