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Real-time PCR-based method for the rapid detection of extended RAS mutations using bridged nucleic acids in
Takao Iida1, Yukie Mizuno1, Yasuharu Kaizaki2
1Department of Clinical Laboratory, Fukui Prefectural Hospital, 2-8-1, Yotsui, Fukui-shi, Fukui 910-8526, Japan.
Abstract:
Mutations in RAS and BRAF are predictors of the efficacy of anti-epidermal growth factor receptor (EGFR) therapy in patients with metastatic colorectal cancer (mCRC). Therefore, simple, rapid, cost-effective methods to detect these mutations in the clinical setting are greatly needed. In the present study, we evaluated BNA Real-time PCR Mutation Detection Kit Extended RAS (BNA Real-time PCR), a real-time PCR method that uses bridged nucleic acid clamping technology to rapidly detect mutations in RAS exons 2-4 and BRAF exon 15. Genomic DNA was extracted from 54 formalin-fixed paraffin-embedded (FFPE) tissue samples obtained from mCRC patients. Among the 54 FFPE samples, BNA Real-time PCR detected 21 RAS mutations (38.9%) and 5 BRAF mutations (9.3%), and the reference assay (KRAS Mutation Detection Kit and MEBGEN™ RASKET KIT) detected 22 RAS mutations (40.7%). The concordance rate of detected RAS mutations between the BNA Real-time PCR assay and the reference assays was 98.2% (53/54). The BNA Real-time PCR assay proved to be a more simple, rapid, and cost-effective method for detecting KRAS and RAS mutations compared with existing assays. These findings suggest that BNA Real-time PCR is a valuable tool for predicting the efficacy of early anti-EGFR therapy in mCRC patients.
Insights
A new real-time PCR method effectively detects RAS and BRAF mutations in metastatic colorectal cancer patients. This simple, rapid, and cost-effective assay aids in predicting anti-EGFR therapy efficacy.
Area of Science:
- Oncology
- Molecular Diagnostics
- Genetics
Background:
- RAS and BRAF mutations predict anti-EGFR therapy response in metastatic colorectal cancer (mCRC).
- Current mutation detection methods may not be sufficiently simple, rapid, or cost-effective for routine clinical use.
- There is a clinical need for efficient molecular diagnostic tools in mCRC patient management.
Purpose of the Study:
- To evaluate the BNA Real-time PCR Mutation Detection Kit Extended RAS for detecting RAS and BRAF mutations.
- To assess the simplicity, rapidity, and cost-effectiveness of the BNA Real-time PCR assay compared to existing methods.
- To determine the utility of BNA Real-time PCR for predicting anti-EGFR therapy efficacy in mCRC.
Main Methods:
- Real-time PCR using bridged nucleic acid (BNA) clamping technology.
- Analysis of 54 formalin-fixed paraffin-embedded (FFPE) tissue samples from mCRC patients.
- Comparison with established KRAS mutation detection kits as reference assays.
Main Results:
- BNA Real-time PCR detected 38.9% RAS mutations and 9.3% BRAF mutations.
- Reference assays detected 40.7% RAS mutations.
- High concordance (98.2%) was observed between BNA Real-time PCR and reference assays for RAS mutations.
Conclusions:
- BNA Real-time PCR is a simple, rapid, and cost-effective method for detecting RAS and BRAF mutations in mCRC.
- The assay demonstrates high accuracy and concordance with existing methods.
- BNA Real-time PCR is a valuable tool for guiding anti-EGFR therapy decisions in mCRC patients.
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