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KRAS mutation detection in colorectal cancer by a commercially available gene chip array compares well with Sanger
Deborah French1, Andrew Smith, Martin P Powers
1Department of Laboratory Medicine, University of California San Francisco, San Francisco, CA 94107, USA. deborah.french@ucsf.edu
Background:
Binding of a ligand to the epidermal growth factor receptor (EGFR) stimulates various intracellular signaling pathways resulting in cell cycle progression, proliferation, angiogenesis and apoptosis inhibition. KRAS is involved in signaling pathways including RAF/MAPK and PI3K and mutations in this gene result in constitutive activation of these pathways, independent of EGFR activation. Seven mutations in codons 12 and 13 of KRAS comprise around 95% of the observed human mutations, rendering monoclonal antibodies against EGFR (e.g. cetuximab and panitumumab) useless in treatment of colorectal cancer.
Methods:
KRAS mutation testing by two different methodologies was compared; Sanger sequencing and AutoGenomics INFINITI® assay, on DNA extracted from colorectal cancers.
Results:
Out of 29 colorectal tumor samples tested, 28 were concordant between the two methodologies for the KRAS mutations that were detected in both assays with the INFINITI® assay detecting a mutation in one sample that was indeterminate by Sanger sequencing and a third methodology; single nucleotide primer extension.
Conclusions:
This study indicates the utility of the AutoGenomics INFINITI® methodology in a clinical laboratory setting where technical expertise or access to equipment for DNA sequencing does not exist.
Insights
KRAS mutation testing in colorectal cancer using the AutoGenomics INFINITI® assay showed high concordance with Sanger sequencing. This assay is a viable alternative for clinical labs lacking advanced sequencing equipment.
Area of Science:
- Oncology
- Molecular Diagnostics
- Genetics
Background:
- Epidermal growth factor receptor (EGFR) signaling pathways regulate cell growth; mutations in KRAS can lead to constitutive pathway activation, independent of EGFR.
- Specific KRAS mutations (codons 12 and 13) are common in colorectal cancer and can confer resistance to EGFR-targeted therapies like cetuximab.
- Accurate KRAS mutation detection is crucial for guiding colorectal cancer treatment decisions.
Purpose of the Study:
- To compare the performance of the AutoGenomics INFINITI® assay against Sanger sequencing for KRAS mutation detection in colorectal cancer.
- To evaluate the clinical utility of the INFINITI® assay in resource-limited laboratory settings.
Main Methods:
- DNA was extracted from 29 colorectal tumor samples.
- KRAS mutation status was determined using both Sanger sequencing and the AutoGenomics INFINITI® assay.
- Results were compared for concordance between the two methodologies.
Main Results:
- High concordance was observed between the INFINITI® assay and Sanger sequencing for detected KRAS mutations.
- The INFINITI® assay identified a mutation in one sample that was indeterminate by Sanger sequencing and single nucleotide primer extension.
- The AutoGenomics INFINITI® assay demonstrated robust performance in detecting common KRAS mutations.
Conclusions:
- The AutoGenomics INFINITI® assay is a reliable method for KRAS mutation testing in colorectal cancer.
- This methodology offers a practical solution for clinical laboratories with limited access to DNA sequencing equipment or technical expertise.
- The INFINITI® assay can aid in selecting appropriate therapies for colorectal cancer patients by accurately identifying KRAS mutations.
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