title: A multiplex SNaPshot assay as a rapid method for detecting KRAS and BRAF mutations in advanced colorectal cancers.
abstract: The analysis of KRAS mutations has become a prerequisite for anti-epidermal growth factor receptor therapy in patients with metastatic colorectal cancers. KRAS mutations are associated with resistance to treatment by monoclonal antibodies such as cetuximab and panitumumab and thus are correlated with a shorter progression-free survival. BRAF mutations also may play a role in treatment decisions. The widespread use of these targeted therapies has generated the need to develop cost-effective methods for routine KRAS and BRAF analysis. The aim of this study was to compare a multiplex SNaPshot assay with DNA sequencing and high-resolution melting analysis for identifying KRAS codons 12 and 13 and BRAF codon 600 mutations. Thus 110 routinely formalin-fixed and paraffin-embedded tissue blocks were tested by each method. The SNaPshot analysis detected KRAS and BRAF codon 600 mutations in, respectively, 34.5% (n = 38) and 10% (n = 11) of these tissue blocks. These results were confirmed by direct DNA sequencing and by high-resolution melting analysis. The costs and time constraints of each detection method were compared at the same time. In conclusion, our newly designed multiplex SNaPshot assay is a fast, inexpensive, sensitive, and robust technique for molecular diagnostic practices and patient selection.
authors: Platform of Molecular Biology of Cancers, University Hospital of Besançon, Besançon, France.
authors: Sandrine
authors: S
authors: Magnin
authors: Erika
authors: E
authors: Viel
authors: Alice
authors: A
authors: Baraquin
authors: Severine
authors: S
authors: Valmary-Degano
authors: Bernadette
authors: B
authors: Kantelip
authors: Jean-Luc
authors: JL
authors: Pretet
authors: Christiane
authors: C
authors: Mougin
authors: Marthe
authors: M
authors: Bigand
authors: Benoît
authors: B
authors: Girardo
authors: Christophe
authors: C
authors: Borg
authors: Christophe
authors: C
authors: Ferrand
mesh_terms: D015179:Colorectal Neoplasms / Q000191:economics / Q000235:genetics* / Q000473:pathology*; D003365:Costs and Cost Analysis; D053938:DNA Fragmentation; D004252:DNA Mutational Analysis / Q000191:economics / Q000379:methods*; D005091:Exons / Q000235:genetics; D006801:Humans; D025202:Molecular Diagnostic Techniques / Q000191:economics; D009154:Mutation / Q000235:genetics*; D009367:Neoplasm Staging; D009691:Nucleic Acid Denaturation / Q000235:genetics; D016133:Polymerase Chain Reaction; D020641:Polymorphism, Single Nucleotide / Q000235:genetics*; D011518:Proto-Oncogene Proteins / Q000235:genetics*; D048493:Proto-Oncogene Proteins B-raf / Q000235:genetics*; D016283:Proto-Oncogene Proteins p21(ras); D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D013997:Time Factors; D018631:ras Proteins / Q000235:genetics*
chemical_list: C117307:KRAS protein, human; D011518:Proto-Oncogene Proteins; C482119:BRAF protein, human; D048493:Proto-Oncogene Proteins B-raf; D016283:Proto-Oncogene Proteins p21(ras); D018631:ras Proteins