Medizinische Universität Graz Austria/Österreich - Forschungsportal - Medical University of Graz
Gewählte Publikation:
SHR Neuro Krebs Kardio Lipid Stoffw Microb
Buxhofer-Ausch, V; Ausch, C; Zeillinger, R; Oberkanins, C; Dandachi, N; Reiner-Concin, A; Kriegshäuser, G.
Duplex reverse-hybridization assay for the simultaneous detection of KRAS/BRAF mutations in FFPE-extracted genomic DNA from colorectal cancer specimens.
Dis Markers. 2013; 34(3):171-177
Doi: 10.3233/DMA-120960
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- Führende Autor*innen der Med Uni Graz
- Kriegshäuser Gernot
- Co-Autor*innen der Med Uni Graz
- Dandachi Nadia
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- Abstract:
- We report the performance evaluation of a non-quantitative reverse-hybridization assay (KRAS-BRAF StripAssay) designed for the simultaneous detection of 10 mutations in codons 12 and 13 of the KRAS gene and BRAF mutation V600E. Dilution experiments using DNA from tumor cell lines or from formalin-fixed paraffin-embedded (FFPE) colorectal cancer (CRC) tissue were performed to assess assay sensitivity. Using 50 ng of total DNA (mutant and wild-type), the KRAS-BRAF StripAssay demonstrated a detection limit of 1% mutant sequence in a background of wild-type DNA. With respect to BRAF V600E, the KRAS-BRAF StripAssay was evaluated using 60 FFPE CRC samples previously analyzed by high resolution melting (HRM). Test strip hybridization identified 2/60 (3%) samples to carry the BRAF V600E mutation, and results were in agreement with those obtained by HRM analysis. This work demonstrates the KRAS-BRAF StripAssay to be a robust and sensitive method for the detection of common KRAS/BRAF mutations in genomic DNA isolated from FFPE tissue samples.
- Find related publications in this database (using NLM MeSH Indexing)
- Adult -
- Aged -
- Aged, 80 and over -
- Colorectal Neoplasms - genetics
- DNA, Neoplasm - analysis DNA, Neoplasm - genetics
- Formaldehyde - chemistry
- Humans -
- Middle Aged -
- Mutation - genetics
- Neoplasm Staging -
- Nucleic Acid Hybridization -
- Paraffin Embedding -
- Proto-Oncogene Proteins - genetics
- Proto-Oncogene Proteins B-raf - genetics
- ras Proteins - genetics
- Find related publications in this database (Keywords)
- KRAS
- BRAF
- mutation detection
- reverse-hybridization
- high-resolution melting