Related Experiment Video
Updated: May 31, 2026
A Multiplexed Luciferase-based Screening Platform for Interrogating Cancer-associated Signal Transduction in Cultured Cells
Published on: July 3, 2013
A multiplex SNaPshot assay as a rapid method for detecting KRAS and BRAF mutations in advanced colorectal cancers
Sandrine Magnin1, Erika Viel, Alice Baraquin
1Platform of Molecular Biology of Cancers, University Hospital of Besançon, Besançon, France.
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.
Insights
A new multiplex SNaPshot assay efficiently detects KRAS and BRAF mutations in metastatic colorectal cancer patients. This cost-effective method aids in selecting patients for targeted therapies like anti-EGFR treatment.
Area of Science:
- Molecular diagnostics
- Oncology
- Genetics
Background:
- KRAS and BRAF mutations are critical biomarkers for anti-EGFR therapy in metastatic colorectal cancer (mCRC).
- These mutations influence treatment response to monoclonal antibodies (e.g., cetuximab, panitumumab), impacting progression-free survival.
- There is a growing need for cost-effective and routine molecular diagnostic methods for KRAS and BRAF analysis in mCRC.
Purpose of the Study:
- To evaluate a novel multiplex SNaPshot assay for identifying KRAS codons 12/13 and BRAF codon 600 mutations.
- To compare the SNaPshot assay's performance against established methods like DNA sequencing and high-resolution melting analysis.
- To assess the cost-effectiveness and time efficiency of the SNaPshot assay for clinical application.
Main Methods:
- A multiplex SNaPshot assay was developed and validated.
- 110 formalin-fixed, paraffin-embedded (FFPE) tissue blocks from mCRC patients were analyzed.
- Each sample was tested using the SNaPshot assay, direct DNA sequencing, and high-resolution melting analysis.
Main Results:
- The SNaPshot assay identified KRAS mutations in 34.5% (n=38) and BRAF V600 mutations in 10% (n=11) of the tested FFPE samples.
- Results from the SNaPshot assay were consistent with those obtained by direct DNA sequencing and high-resolution melting analysis.
- Comparative analysis indicated the SNaPshot assay offers advantages in terms of cost and time.
Conclusions:
- The developed multiplex SNaPshot assay is a sensitive, robust, fast, and inexpensive method for detecting KRAS and BRAF mutations.
- This assay is suitable for routine molecular diagnostic practices and patient selection for targeted therapies in mCRC.
- The SNaPshot assay provides a valuable tool for optimizing anti-EGFR treatment strategies.
More Related Videos
08:23Single Droplet Digital Polymerase Chain Reaction for Comprehensive and Simultaneous Detection of Mutations in Hotspot Regions
Published on: September 25, 2018
10:16Employing Digital Droplet PCR to Detect BRAF V600E Mutations in Formalin-fixed Paraffin-embedded Reference Standard Cell Lines
Published on: October 8, 2015